WAKO(和光纯药)介绍

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WAKO 和光纯药介绍

WAKO 和光纯药介绍代理商上海金畔生物,欢迎广大新老客户访问日本Wako试剂官网或者咨询我们获取更多详细信息。

日本和光纯药试剂

Wako和光纯药官网: http://www.wako-chem.co.jp
备 货 期: 部分现货或2-3周

Wako(和光纯药)来自日本,是日本最大的实验室化学试剂供应商,Wako是世界五大实验室化学试剂供应商之一,是全球第一流的试剂制造厂商,以其高品质的试剂闻名世界。Wako”系列试剂涵盖了全面的生物化学、分析化学、有机化学、环境分析、食品.医药品分析、高纯度及认证标准品等多类,约有50万种试剂。在基因组学、蛋白质组学研究产品开发方面Wako同样强而有力。Wako也是继承了日本企业一贯的严谨品质,获得了ISO9001质量管理体系认证和ISO/IEC17025实验室定。

产品品牌:Wako。

Wako日本和光纯药工业株式会社是全球第一流的试剂制造厂商,成立八十多年一直致力于高质量的试剂及药业的生产与开发,所生产的各类产品均获得 ISO9002 认可。试剂涵盖了全面的生物化学、分析化学、有机化学、环境分析、食品·医药品分析、高纯度及认证标准品等多类,超过六万种试剂。
日本和光纯药工业株式会社(WAKO)生产的各种试剂,主要包括以下几大类:
一、生物化学试剂 1 遗传因子 2 免疫研究试剂 3 血凝、纤溶试剂 4 组织化学研究试剂 5 酶、蛋白质、肽 6 脂类 7 二十碳醇类 8 类固醇、甾体、激素 9 单糖、寡糖、多糖、环化糊精类 10 神经作用物质 11 生理作用物质 12 毒素 13 抗生素、抑制剂 14 膜蛋白分析试剂 15 缓冲剂(good`s buffers) 16 细胞研究试剂 17 细胞培养试剂(1)动物组织培养试剂(2)植物组织培养试剂(3)微生物培养试剂 18 氨基酸分析试剂 19 电泳试剂 20 其它生化试剂

二、分析试剂 1 层析试剂(1)高效液相色谱试剂(2)萃取试剂(3)柱层析试剂(4)薄层层析试剂(5)气相色谱试剂 2 原子吸收分析试剂 3 吸光分析试剂 4 紫外分析试剂 5 红外分析试剂 6 化学发光分析试剂 7 荧光分析试剂 8 LC/MS试剂 9 NMR 试剂 10 ESR 试剂 11 辐射测定试剂 12 元素分析试剂 13 滴定用试剂 14 指示剂 15 标准缓冲液 16 PH 标准试剂 17 电极试剂 18 容量分析试剂 19 精密分析试剂 20 超微量分析试剂 21 高纯试剂

三、环境分析用试剂 1 环境激素分析试剂 2 农药残留检测试剂 3 二恶英测定试剂 4 水质分析试剂 5 大气污染物质测定试剂 6 食品、药品分析试剂 7 LAL test 仪器、试剂 8 变异原(突变诱导)检测试剂 9 细胞繁殖、毒性研究试剂 10 家庭用品安全检测试剂 11 合成洗涤剂检测试剂(衣料用) 12 石油及制品检测试剂

四、合成用试剂 1 核酸合成试剂 2 多肽合成试剂 3 有机合成溶剂 4 Grignard 格氏反应试剂 5 氨基化试剂 6 重氮烷基化试剂 7 卤化试剂 8 金属触媒 9 光学活性物质 10 光学拆分试剂 11 半合成催化剂 12 绿色化学关联试剂(微胶囊化催化剂、树脂基质两性分子磷化钯催化剂)

五、材料研究用试剂 1 高纯度化合物 2 功能性新材料

六、实验室用清洗剂 1 实验室用洗涤剂 2 其它洗涤剂

七、临床诊断试剂

分类浏览:

所有分类 生命科学 (1) 二恶英检测 (1) Wako日立氨氨基酸分析 (52) 固相萃取柱 (140) 三聚氰胺 (4) 苏丹红 (4) 食品禁用色素 (4) 维生素 (13) 甜菊叶提取物 (18) 表面润湿张力 (36) 高血压研究 (10) 抑制剂 (24) 标签蛋白抗体 (62) 细胞因子 (268) 生理活性物质 (3) DNA marker (22) 微生物与细胞培养 (32) 干细胞培养 (78) SU受体选择 (3) Wako多聚蛋白胨系列 (13)  Wako Phostag磷酸化蛋白解析应用 (23) 成像系统发光试剂 (6) 发光试剂 (16) 活性氧测定用化学发光探针 (5) 生物影像用荧光试剂 (30) 组织透明化辅助试剂 (23) 老年痴呆/阿尔茨海默症研究 (56) 癌症研究 (99) 发育与再生神经研究 (3) 肥胖研究 (7) 高胆固醇血症研究 (15) 聚糖/寡糖 (1) 蛋白质及酶 (1) 生物染料和指示剂 (1) 通用试剂 (5) 骨代谢研究 (5) 骨骼疾病研究 (1) 精神分裂症的研究 (19) 抗炎研究 (2) 脑损伤研究 (3) 脑肿瘤和癌症研究 (6) 帕金森病研究 (1) 神经病理性疼痛的研究 (1) 神经系统细胞因子 (21) 神经系统信号研究 (2) 肾脏研究 (1) 睡眠研究、摄食研究 (3) 糖尿病、肥胖的研究 (44) 心血管研究 (2) 嗅觉神经研究 (1) 血管生成促进剂 (2) 氧化磷酸化抑制剂 (98) 氧化应激研究 (2) 抑郁症研究 (38) 其它 (8) 类视黄醇,视黄酸 (8)岩藻黄素 (4) 黄酮类 (26) 茶多酚 (24) 大豆苷 (21) SILAC (26) 抗生物素蛋白 (13) CultureSure 氨基酸系列 (42) 动物细胞培养 (97) 胶原酶系列 (15) 昆虫细胞培养 (3) 代理品牌 (946) 缓冲液 (39) 显色剂 (8) Wako氨基酸标准品 (8) 再生液 (7) 氨基酸序列分析 (27) 日本药典草药测试用 (64) 草药测试用标准品 (2) 法定薄层色谱专用草药 (2) 生物化学测试专用草药 (72)

Wako(和光纯药)公司产品标签变更通知

和光纯药工业株式会社一直以来承蒙各方关照,借此表示衷心的感谢。

和光自2018年4月1日起将公司名变更为下述名称。

新公司名称:富士フイルム和光純薬株式会社

         FUJIFILM Wako Pure Chemical Corporation

*公司地址和电话号码与原来一致。

另外,和光产品标签也会有相应的调整。

由此给您带来不便,我们深表歉意!今后仍望惠顾关照!有关标签的内容,调整如下:

新标签

备注

旧标签

公司名称和标签变更期间的规定

标签

自2018年4月起生产的产品将更换成新标签。

新标签产品与旧标签产品并行销售。

检测报告

2018年4月后发布的检测报告将使用新的公司名称。

SDS

自2018年4月起将陆续更换为新的公司名称,使用当前公司名称的SDS也可继续使用。

包装材料

和光将逐步更换一部分印有当前公司名称和品牌logo的包装材料,旧包装2018年4月后仍会使用。

包装材料的质地没有改变,不会对产品质量造成影响。

Lumiprobe代理(Cy系列活性荧光染料) 金畔生物修饰性PEG(单分散短链小分子PEG)  Laysan bio修饰性PEG代理  NANOC进口PEG代理  Avanti进口修饰性PEG代理  wako日本和光代理(表面张力测试试剂、日立氨基酸分析仪配套试剂、低聚果糖标准品、Anti Iba1, Rabbit抗体)  日本关东化学Kanto试剂代理  Megazyme代理  Hampton代理(蛋白结晶) whatman代理(whatman滤膜、whatman滤纸) GE代理(蛋白纯化柱及填料)   Sigma优势代理 TCI代理 Amresco代理 MPbio代理(动物肠膜炎DSS)Merck Millipore代理 Roche酶代理  NEB酶代理  Abcam代理  CST代理  Santa Cruz代理  Biolegend代理Invitrogen代理   millipore代理  BD流式抗体代理   GeneTe x抗体代理  Novus抗体代理  R&D代理  ebioscience代理  Biovison代理  Jackson代理  MBL抗体代理  ProSpec抗体代理  Bethyl抗体代理  Antibody Revolution抗体代理  Torrey Pines Biolabs代理Toyobo酶代理  TRC标准品代理 USP代理  EP代理  Dr代理 Reagecon代理   Corning康宁(培养皿、培养板)代理  Axygen代理  Falcon耗材代理   Nunc耗材代理  NEST耗材代理  OXOID代理 NISSUI日水培养基代理 Himedia培养基代理 BD培养基代理(灭活结核杆菌231141)  Ludger代理(糖蛋白分析产品)  Eppendorf代理  Bio-Rad伯乐代理  wheaton代理 Labnet代理  Rearch dites、Harlan品牌动物饲料代理 标准品代理 抗体代理 酶试剂代理Elisa试剂盒代理、柔软型灌胃针 小鼠灌胃针、小鼠记号笔、进口动物玩具 动物投药胶囊。

Wako 292-81101生物制药残留DNA提取试剂盒(碘化钠法)

Wako 292-81101生物制药残留DNA提取试剂盒(碘化钠法)

生物制药残留DNA提取试剂盒(碘化钠法)残留DNA提取试剂盒遵照中国药典记载的碘化钠法,适用于疫苗和治疗用生物制品所含宿主细胞残余DNA的提取。

残留DNA提取试剂盒可以提取样品中含有的极微量的DNA,回收率较高。DNA的提取操作时间为60-90 min。提取的DNA可以通过qPCR进行定量。

在工业生产的疫苗和治疗用生物制品(干扰素、白细胞介素、单克隆抗体、重组蛋白)中,宿主细胞残余DNA污染直接影响最终产品和整个生产过程的质量和产量,可能引起动物致瘤、感染病毒DNA等严重后果,中国药典、WHO、EU、FDA对宿主细胞DNA残留限量都有规定。传统的DNA提取方法蛋白酶K-SDS法在SDS的稳定和促进下降解细胞中的蛋白质特别是核酶,再经有机溶剂萃取得到高质量的DNA,但毒DNA和生物制品宿主细胞残余DNA的提取试剂盒。使用新型提取步骤,用碘化钠(NaI)作为促溶剂,稳定且灵敏度高、时间短,无有毒溶剂,无需复杂和费力操作,即可从样品中得到高质量和高回收率的DNA。

◆特点
● 遵照中国药典登载的碘化钠法*1

● 高回收率

● 单管操作(无需换管) ,耗时仅60-90 min

● 不需要苯酚和氯仿等有机溶剂

● 兼容多种DNA

*1 为使DNA检测方法与国际标准保持同步,中国药典(The Chinese Pharmacopoeia)宣布建立残留细胞DNA检测国家标准。碘化钠法将作为残留DNA提取方法,被新增至2020版的中国药典中。

◆原理

1. 使样品中的蛋白质和脂质溶于碘化钠和N-月桂基肌氨酸钠;

2. 异丙醇与糖原选择性地共沉淀DNA。

◆使用方法

方法1

方法2

◆应用实例

DNA spiking test(加标回收实验)

161579152874.png 161579152893.png
161579152967.png
■ 实验方案
方案1:血清中DNA 提取操作步骤
试剂准备:
1) DNA 提取之前,试剂提取如下:
26 ml 碘化钠溶液加入6 ml 蒸馏去离子水稀释,
再加入1 ml 月桂酰肌氨酸钠和65 μl 糖原溶液并混合均匀。
2) 向洗涤溶液(B)加入2 μl 糖原溶液并立即混合均匀。
注意事项:
1) 用于稀释Nal 溶液的蒸馏去离子水须去除DNA。
2) 配制好的洗涤溶液(B)在4℃至少能保存一周。如果需要少量的洗涤溶液(B),加入糖原溶液到无菌离心管中至合适体积即可。
3) 在保存期间Nal 溶液会结晶,加热溶液至50℃可帮助溶解。
DNA 提取步骤:
1) 吸取100 μl 的血清样品加入1.5 ml 微量离心管;
2) 再加入300 μl 配制好的Nal 溶液至离心管并混合均匀;
3) 将离心管在60℃恒温容器中加热15 分钟;
4) 取出离心管,加入400 μl 异丙醇至离心管并混合均匀;
5) 离心管在室温静置15 分钟后离心10,000 g 15 分钟;
6) 尽可能多地吸取上清液,将离心管倒置在纸面上除去残留溶液;
7) 加入1 ml 配制好的洗涤溶液(B)重悬浮得到的白色沉淀,并使白色沉淀从离心管壁上弹开;
8) 短暂离心10,000 g 5 min,去除上清液,将剩余沉淀真空干燥,用于后续的DNA 分析。
 
方案2: Threshold* 系统DNA定量法的预处理
试剂预处理:
1) 将65 μl 糖原溶液加入26 ml 碘化钠溶液;
2) 将2 μl 糖原溶液加入40 ml 洗涤溶液(B),立即混合均匀。
注意事项:
1) 用于稀释碘化钠溶液的蒸馏去离子水须去除DNA。
2) 配制好的洗涤溶液(B)在4℃至少能保存一周。如果需要少量的洗涤溶液(B),加入糖原 溶液到无菌离心管中至合适体积即可。
3) 在保存期间碘化钠溶液会结晶,加热溶液至50℃可帮助溶解。
4) 提取步骤应在DNA 变性之前,因此重要的是使用无菌技术尽可能降低DNA 污染。预先包装好的无菌吸管、无菌试管和手套都要经过此步骤。
DNA 提取步骤:
1) 吸取400 μl 或500 μl 的样品加入2 ml 无菌带盖微量离心管并混合均匀;
2) 吸取20 μl 的月桂酰肌氨酸钠溶液加入微量离心管并混合均匀;
3) 再加入500 μl 包含糖原的碘化钠溶液至混合溶液,涡旋后在40℃保温15 分钟;
4) 加入900 μl 异丙醇至混合溶液,涡旋后静置于室温15 分钟;
5) 短暂离心10,000 g 15 min 后,能看见管壁有白色沉淀,吸取上清液后将离心管倒置于纸面去除残留溶液;
6) 加入800 μl 洗涤溶液(A)至离心管,剧烈涡旋使白色沉淀从离心管壁上弹开;
7) 短暂离心10,000 g 5 min 后,去除残留溶液;
8) 加入1500 μl 包含糖原的洗涤溶液(B)至离心管并涡旋,短暂离心10,000 g 5 min 后吸取上清液,剩余的白色沉淀包括DNA 和糖原载体;
9) 加入分析用缓冲液(Zero Calibrator buffer) 使白色沉淀溶解,用于后续的DNA 分析。

DNA Extractor™ Kit可以高产量提取残留DNA,甚至少量残留DNA。

■ 试剂
1.试剂盒组分:
1)碘化钠溶液  1 x 26 ml
2)月桂酰肌氨酸钠  1 x 1.2 ml
3)洗涤溶液(A)  1 x 42 ml
4)洗涤溶液(B)  2 x 40 ml
5)糖原溶液  1 x 0.1 ml
2.必需的试剂和仪器:
试剂:
1)异丙醇(特级) 40 ml
2)蒸馏去离子水 6 ml
仪器:
1)微型离心机(Max. 12,000 g)
2)有盖微量离心管(1.5 or 2 ml)
3)旋涡混合器
■ 保存
2 – 10℃,试剂盒的保存期为2 – 3 年。

◆试剂盒构成

产品编号 内容物 内容量
299-81111 碘化钠溶液, CP 26 mL
296-81121 N-月桂酰肌氨酸钠溶液, CP 1.2 mL
293-81131 洗净液A, CP 42 mL
290-81141 洗净液B, CP 40 mL
297-81151 糖原溶液, CP 0.1 mL

备注:每种成分不单独出售

◆产品列表

产品编号 产品名称 包装
292-81101 残留DNA提取试剂盒(碘化钠法)
DNA Extractor™ Kit for Residual DNA, CP Method (Sodium Iodide Method)
50 tests
温馨提示:不可用于临床治疗。

 

Wako DNA Extractor®提取试剂盒产品选择
样本 全血 血清 血浆 组织 石蜡包埋组织切片 生物制药 培养细胞 头发、指甲、血迹、唾液
基因组DNA DNA Extractor ® WB Kit
DNA Extractor ® WB-Rapid Kit
DNA Extractor ® WB Kit DNA Extractor ® SP kit
DNA Isolator PS-Rapid Reagent
DNA Extractor ® WB Kit DNA Extractor ® FM kit
线粒体DNA(mt DNA) mtDNA
Extractor ® WB Kit
mtDNA Extractor® WB Kit mtDNA
Extractor ® CT Kit
游离DNA DNA Extractor ® SP kit DNA Extractor ® SP kit
低氧化程度的DNA DNA Extractor ® WB Kit DNA Extractor ® TIS kit DNA Extractor ® TIS kit
细菌基因组DNA DNA Extractor ® kit DNA Extractor ® kit
病毒DNA DNA Extractor ® kit

 

DNA Extractor® 系列产品列表
产品编号 产品名称 规格 样品类型 原理 特点 试剂盒组成
生物制品中的残余DNA提取
295-50201 DNA Extractore® Kit Wako
生物制药残余DNA抽提试剂盒
50次 生物制药
血清
碘化钠法 生物制品中的残余DNA提取 碘化钠溶液
月桂酰肌氨酸钠溶液
洗涤液(A)
洗涤液(B)
糖原溶液
26ml×1
1.2mL×1
42mL×1
40mL×2
0.1mL×1
血清和血浆中的DNA提取
296-60501 DNA Extractor® SP Kit
血清血浆中DNA抽提试剂盒
50次 血清和
血浆
碘化钠法 1、高灵敏度仅需100 μL血清或血浆样品,高DNA产量;
2、整个提取过程在单独的离心管中,无交叉污染;
3、特别配制的乙醇能完全去除血液中的脂质
酶反应液
蛋白质消化液
碘化钠溶液
乙醇溶液
洗涤液(A)
洗涤液(B)
10ml×1
25μL×1
15mL×1
30mL×1
50mL×1
50mL×1
人和动物组织中的DNA提取(用于8-羟基脱氧鸟苷检测)
296-67701 DNA Extractor® TIS Kit
人和动物薄壁组织中尿8-羟基脱氧鸟苷DNA抽提试剂盒
50次 组织培养
细胞
碘化钠法 1、用于8-羟基脱氧鸟苷(氧化应激标志物)检测和鉴定;
2、抗氧化剂阻止更进一步的DNA氧化
裂解液
酶反应液
蛋白酶溶液
蛋白消化液
氧化抑制剂
碘化钠溶液
乙醇溶液
PEG溶液
75mL×2
15ml×1
50μL×1
750μL×1
350μL×1
15mL×1
30mL×1
20mL×1
全血中的DNA提取
291-50502 DNA Extractor® WB Kit
全血DNA抽提试剂盒
50次 全血
组织
培养细胞
碘化钠法 1、高DNA回收率;
2、整个提取过程在单独的离心管中,无交叉污染
裂解液
酶反应液
碘化钠溶液
洗涤液(A)
洗涤液(B)
蛋白酶
65ml×2
10mL×1
15mL×1
50mL×1
50mL×1
10mg×1
297-54801
293-54803
DNA Extractor® WB-Rapid Kit
全血DNA抽提试剂盒
20次
200次
全血 比DNA Extractor® WB Kit实验步骤更少、提取时间更短(30分钟以内)。 裂解液
酶反应液
洗涤液
蛋白酶溶液
20次反应
10mL×1
0.8mL×1
20mL×1
40μL×1
200次反应
50mL×1
8mL×1
70mL×3
400μL×1
石蜡包埋组织的DNA提取
295-52401 DNA Isolator PS Kit
石蜡包埋组织DNA抽提试剂盒
100次 石蜡包埋
组织
酶反应液
酶激活剂
蛋白酶
DNA沉淀促进剂
DNA稀释剂
2ml×1
34mg×1
2.2mg×1
0.5mL×1
0.5mL×1
291-56401 DNA Isolator PS-Rapid Reagent
石蜡包埋组织DNA抽提试剂盒
100次 石蜡包埋
组织
煮沸法 DNA提取溶液 10mL×5
全血中线粒体DNA提取
293-54401 mtDNA Extractor® WB Kit
全血中线粒体DNA抽提试剂盒
25次 全血 碘化钠法 全血中线粒体 DNA提取可在90分钟内完成 白血球提取溶液
DNA提取溶液|
DNA提取溶液||(A)
DNA提取溶液||(B)
DNA提取溶液|||
碘化钠溶液
洗涤液
5.0ml×1
26.5ml×1
1.3ml×1
1.3ml×1
1.9mL×1
7.5mL×1
50mL×1
培养细胞和组织样品中的线粒体DNA提取
291-55301 mtDNA Extractor® CT Kit 25次 组织
培养细胞
碘化钠法 培养细胞和组织样品中的线粒体
DNA提取
匀浆液援冲液
DNA提取溶液|
DNA提取溶液||(A)
DNA提取溶液||(B)
DNA提取溶液|||
碘化钠溶液
洗涤液
25.0ml×1
1.3ml×1
1.3ml×1
1.3ml×1
1.9mL×1
7.5mL×1
50mL×1
法医学调查的DNA提取
295-58501 DNA Extractor® FM Kit
法医学研究DNA抽提试剂盒
50次 头发
指甲
血迹
唾液
碘化钠法 在几十分钟内可溶解多种毛发 裂解液
酶激活试剂(EAR)
再水化液
蛋白酶
碘化钠溶液
洗涤液(A)
洗涤液(B)
9.5ml×1
80mg×1
0.5ml×1
10mg×1
12.5mL×1
50mL×1
50mL×1
上述试剂仅供实验研究用,不可用作“医药品”、“食品”、“临床诊断”等。

Wako 残留DNA提取试剂盒 DNA with DNA Extractor® Kit

Wako 残留DNA提取试剂盒 DNA with DNA Extractor® Kit

This article was written by Hiroki Fukuchi, Life Science Research Laboratories of FUJIFILM Wako Pure Chemical Corporation, Japan.

导言
因为许多生物药物,包括疫苗,都是由培养的细胞或微生物(如大肠杆菌指出宿主细胞来源的DNA可能存在于合成的药物物质和产物中.鉴于残留DNA可能转移宿主细胞或病毒衍生的癌基因,病毒DNA可能导致感染事件,定量检测残留DNA是制造业的重要测试,也是生物制药过程验证等测试的一部分。正如一份报告所指出的那样,每一剂量的残余DNA最多可达100 pg,这对于生物制药来说是可以接受的,1)不仅在美国和欧洲国家,而且在其他国家,越来越需要对宿主细胞中的残留DNA进行定量检测。为了检测残留DNA的痕迹,必须从样品中提取出高收率的残留DNA。本文旨在介绍DNA提取器的用途。®用于检测残留DNA作为DNA纯化试剂的试剂盒。

DNA提取器®试剂盒(#295-50201,Fujifilm Wako纯化学公司)
在对生物制品中的总DNA进行检测和定量之前,有必要从蛋白质等其他生物成分中分离纯化生物制品中的DNA。一般来说,DNA的分离是通过蛋白酶介导的样品消化,然后用苯酚和/或氯仿提取的。然而,这种方法的缺点是,必须使用有害的苯酚和氯仿,需要时间和人力的提取,虽然可以获得相当高纯度的DNA。此外,二氧化硅载体固相萃取,由于载体吸附导致DNA丢失,对DNA的提取效果不理想。有机溶剂的提取也不利于DNA的有效回收,这是DNA提取试剂需要解决的关键问题之一。

DNA提取器®试剂盒(产品代码:295-50201),我们于1992年推出,解决了上述限制,使用钠碘提取高纯度的DNA,通过简单的程序。

DNA提取原理®试剂盒
工具包的组成部分*含有碘化钠和表面活性剂,当2-丙醇加入时,它们通过溶解蛋白质和其他生物成分,选择性地沉淀(共沉淀)核酸(主要是DNA)和糖原,作为蛋白质增溶剂(潮致离子)。2)与上述方法不同,纯化步骤被简化为在没有载体或有机溶剂的情况下产生沉淀物,从而能够以较高的收率提取微量DNA。

*工具包的组成部分:
碘化钠溶液,N-月桂酸盐溶液,洗涤液(A),
洗涤液(B),糖原溶液

DNA提取器提取DNA的实例®试剂盒和总DNA定量
本文介绍了一份关于在稀释剂或辅料存在的情况下DNA产量的报告,该报告最初发表在瓦科纯化学杂志第60卷第3期(1992年)第28页。本实验通过在磷酸盐缓冲液中溶解常用作稀释剂或辅料(如精氨酸、尿素)或蛋白质(BSA和人γ-球蛋白)的常用或过量的物质制成模型溶液,并在每个模型溶液中加入一定量的小牛胸腺脱氧核糖核酸(PG)。然后,根据dna提取器标签上的说明,从模型溶液的400μL中提取dna。®基特。所得沉淀物溶于500μL磷酸盐缓冲液中,用阈值法测定总dna。®总DNA分析系统,*3,4)来测量DNA的产量。测量结果见表1。

Introduction

Since many biopharmaceuticals, including vaccines, are manufactured from cultured cells or microorganisms such as Escherichia coli, it is pointed out that host cell-derived DNA may remain in resultant drug substances and products. Given the possibility that the residual DNA may transfer host cell- or virus-derived oncogenes and that viral DNA may cause infectious events, quantitative detection of residual DNA is important testing for manufacturing and as part of testing such as process validation for biopharmaceuticals. As indicated by a report recommending that up to 100 pg of residual DNA per dose is acceptable for biopharmaceuticals,1) there is increasing need for quantitative detection of residual DNA from host cells as a quality test not only in the US and European countries but also in other countries. To detect traces of residual DNA, it is necessary to extract traces of residual DNA from a sample at a high yield. This article is intended to introduce the usefulness of DNA Extractor® Kit in detecting traces of residual DNA as a DNA purification reagent.

DNA Extractor® Kit (#295-50201, FUJIFILM Wako Pure Chemical Corporation)

Prior to detection and quantification of total DNA in a biologics, it is necessary to separate and purify DNA in the biologics from other biological components such as proteins. In general, DNA is isolated through protease-mediated digestion of specimen followed by extraction with phenol and/or chloroform. However, this method has disadvantages that deleterious phenol and chloroform have to be used and that time- and labor-consuming extraction is required, although quite highly pure DNA can be obtained. In addition, solid-phase extraction via silica carriers, which results in loss of DNA due to adsorption on carriers, is not ideal to recover traces of DNA. Extraction with organic solvents is also not favorable with inefficient recovery of traces of DNA, which is a key issue to be addressed with DNA extraction reagents.

DNA Extractor® Kit (Product code: 295-50201), which we launched in 1992, resolves the aforementioned limitations by using sodium iodide to extract highly pure DNA at high yields through simple procedures.

Principle of DNA Extractor® Kit

The components of the kit* contain sodium iodide and surfactants, which act as protein solubilizers (chaotropic ions) by solubilizing proteins and other biological components in specimens to precipitate (coprecipitate) nucleic acids (mainly DNA) and glycogen selectively when 2-propanol is added.2) Purification steps are simplified to produce precipitates without carriers or organic solvents, unlike the aforementioned methods, enabling the extraction of traces of DNA at high yields.

* Components of the kit:
Sodium Iodide Solution, Sodium N-Lauryl Sarcosinate Solution, Washing Solution (A),
Washing Solution (B), Glycogen Solution

Example of DNA extraction with DNA Extractor® Kit and total DNA quantification

A report on yields of DNA in the presence of diluents or excipients with the use of the kit, which was originally published on page 28 in Wako Pure Chemical Jiho Vol. 60 No. 3 (1992), is introduced here. In this experiment, model solutions were prepared by dissolving usual or excessive doses of substances often used as diluents or excipients (e.g., arginine, urea) or proteins (BSA and human γ-globulin) in phosphate-buffered saline, and a given amount (pg) of calf thymus DNA was added to each model solution. Subsequently, DNA was extracted from 400 μL of model solution according to the instructions on the label of DNA Extractor® Kit. The obtained precipitates were dissolved in 500 μL of phosphate-buffered saline, and total DNA was quantified using Threshold® Total DNA assay system,*3,4) to measure the yield of DNA. Measurement results are presented in Table 1.

Table 1. Yields of DNA in the presence of diluents or excipients
Diluent and its amount Amount of DNA (pg) Yield of DNA (%)
Sorbitol 200 mg/mL
50 mg/mL
50
50
95
94
Maltose 400 mg/mL
200 mg/mL
50 mg/mL
50
50
50
89
102
98
Mannitol 200 mg/mL 50 84
Dextrose 200 mg/mL 50 81
Sucrose 200 mg/mL 50 92
Urea 1 mol/L 50 106
Arginine 200 mg/mL 50 110
γ-globulin 60 mg/mL
60 mg/mL
60 mg/mL
10
5
2.5
102
84
88
BSA 200 mg/mL 10 95

For all five sugars at one or more concentrations, 80% to 110% of DNA was recovered. For arginine and urea, DNA was extracted at a high yield. For both BSA and human γ-globulin, the proteins tested, the yield of DNA was high (some proteins in specimens may precipitate, but the precipitation can be managed with dilution or proteolytic enzymes5)). The aforementioned results show that the kit can extract residual DNA from various samples at high yields in a highly reproducible manner.

* Threshold® Total DNA assay system is manufactured by Molecular Devices for total DNA quantification and designed to quantitatively measure DNA as a molecule rather than as a gene. This system has a detection sensitivity of 2 pg/assay for total DNA.

Example of extraction of traces of residual DNA with DNA Extractor® Kit and DNA quantification by qPCR assay

As mentioned above, Threshold® Total DNA assay system is intended to quantify total DNA, but not to detect any specific gene. Considering the advantages of qPCR quantification for detecting a specific gene, which can be used to detect residual DNA clearly derived from host cells more sensitively than total DNA quantification with Threshold® Total DNA assay system, it was investigated whether traces of DNA could be extracted from potential host cells.

In this experiment, genomic DNA from CHO cells or Escherichia coli, which are widely used to produce proteins and antibodies, was used as a residual DNA model to extract and quantify traces of residual DNA using a qPCR assay. The experimental procedure is presented in Figure 1.

Figure 1. Experimental procedure

wb021698_img01.png

First, DNA was extracted from samples spiked with 0.1 to 100 pg of genomic DNA in 100 μL of water (distilled water for injection) using the kit and then dissolved in 100 μL of water. Subsequently, a qPCR assay was performed to calculate the yield of DNA from a calibration curve prepared simultaneously. The yield of DNA was 85% to 120% for Escherichia coligenome in the range of 1 to 100 pg and for CHO cell genome in the range of 0.1 to 100 pg (the yield was almost 100% up to 1,000 pg for both genomes, although no data are presented here).

Another experiment was performed based on the assumption that residual DNA was contained in a cell culture supernatant. After Panc-1, a human pancreatic cancer-derived cell line, was cultured in 10% FBS DMEM for 3 days, 0.1 pg of genomic DNA from CHO cells was added to 500 μL of culture supernatant to prepare a cell culture supernatant sample. The yield was calculated to be 0.093 pg from a calibration curve. This experiment showed that 0.1 pg (100 fg) of residual DNA in a 500-μL sample, as small as trace in fg units, can be extracted at a high yield using the kit. In addition, traces of residual DNA was recovered from all samples, including water, phosphate-buffered saline, and cell culture supernatant, at as high yields as almost 90% or more (Tables 1 and 2), suggesting that the kit can extract traces of residual DNA from various samples at high yields, as mentioned in the section of total DNA quantification.

Table 2. Yields of genomic DNA
Sample
(added genomic DNA)
Amount of DNA Amount of recovered DNA determined from calibration curve Yield
Distilled water for injection
(Escherichia coli)
0.1pg ND (less than lower limit of detection)
1pg 1.031pg 103%
10pg 11.09pg 111%
100pg 85.1pg 85%
Distilled water for injection
(CHO cells)
0.1pg 0.933pg 93%
1pg 1.187pg 119%
10pg 11.57pg 116%
100pg 87.1pg 87%
Human cell culture supernatant (CHO cells) 0.1pg 0.0928pg 93%

Conclusion

Residual DNA was recovered from samples at high yields using the kit. DNA extraction from samples is a very important step for quantifying residual DNA, and the kit can extract traces of residual DNA at high yields. In addition, since the kit can be used in various samples and may be useful in extracting residual DNA not only from CHO cells but also from other host cells such as Escherichia coli and yeast, the kit is expected to be used for testing of biopharmaceuticals in future.

References  参考文献

  1. Knezevic et al, Biologicals 36:203-211 (2008)
  2. Ishizawa, M et al, Nucleic Acids Res., 19, 5792 (1991)
  3. Kung. V. T et al, Anal. Biochem. 187, 220-227 (1990)
  4. Mizusawa S, Honma R, Pharm Tech. Japan 7, 309-314, 426-431 (1991)
  5. Wako Pure Chemicals Jiho Vol. 61 No. 1 p. 27 (1993)

1.Related product
DNA Extractor® KitRelated Products
DNA Extractor® Kit Series

Wako 聚乙烯醇Poly(vinyl Alcohol CAS 9002-89-5

Wako 聚乙烯醇Poly(vinyl Alcohol  CAS  9002-89-5

WAKO 和光纯药介绍代理商上海金畔生物,欢迎广大新老客户访问日本Wako试剂官网或者咨询我们获取更多详细信息。

品牌 产品编号 产品名称 等级 规格 CAS No.
Wako 163-03045 Polyvinyl Alcohol (Polymerization Degree about 500) 聚乙烯醇(聚合度 500) No grade 500 g 9002-89-5
Wako 165-16315 Poly(vinyl Alcohol) 500, Completely Hydrolyzed  聚乙烯醇 500,完全水解 Wako 1st Grade 500 g 9002-89-5
Wako 165-17915 Polyvinyl Alcohol 聚乙烯醇 JIS Special Grade 500 g 9002-89-5
Wako 160-08295 Polyvinyl Alcohol  聚乙烯醇 for Absorptiometric Analysis 500 g 9002-89-5
Wako 160-03055 Polyvinyl Alcohol (Polymerization Degree about 1500) 聚乙烯醇(聚合度约1500) No grade 500 g 9002-89-5
Wako 160-11485 Polyvinyl Alcohol 聚乙烯醇 Wako 1st Grade 500 g 9002-89-5
Wako 162-16325 Poly(vinyl Alcohol) 1,000, Completely Hydrolyzed 聚乙烯醇 1000,完全氢化水解 Wako 1st Grade 500 g 9002-89-5
Wako 163-16355 Poly(vinyl Alcohol) 3,500, Partially Hydrolyzed 聚乙烯醇 3500,部分水解 Wako 1st Grade 500 g 9002-89-5
Wako 167-03065 Polyvinyl Alcohol (Polymerization Degree about 2000) 聚乙烯醇(聚合度 2000) No grade 500 g 9002-89-5
Wako 169-16335 Poly(vinyl Alcohol) 1,000, Partially Hydrolyzed 聚乙烯醇 1000,部分水解 Wako 1st Grade 500 g 9002-89-5

Wako 和光纯药 011-27991 Anti Iba1, Goat Iba1抗体,山羊源多克隆抗体说明书

Wako 和光纯药 011-27991 Anti Iba1, Goat  Iba1抗体,山羊源多克隆抗体说明书

【Background】
Iba1 is a protein highly expressed in microglia and macrophages with a molecular weight of about 16.7 kDa1). The protein is a commonly known microglial marker in the nervous system.This item is a goat polyclonal antibody that reacts with Iba1.
For Research Use Only. Not for use in diagnostic procedures or therapeutic use.

小胶质细胞标记
Iba1是一种约17 kDa的蛋白,在神经系统小胶质细胞中特异性表达,经常被用作小胶质细胞标记物。 本产品是识别Iba1的山羊多克隆抗体。

【Description】

[Purification] Purified from the goat serum by antigen-affinity chromatography
[Reactivity] Reacts with Iba1
[Antigen] Synthetic peptide corresponding to the C-terminus of Iba1
[Clone No.] -( polyclonal)
[Species cross reactivity] Mouse and Rat(Other species have not been tested)
[Host] Goat
[Concentration] indicated on the label
[Formulation] TBS

[纯化]通过抗原亲和层析从山羊血清中纯化
[反应性]与Iba1反应
[抗原]对应于Iba1 C末端的合成肽
[克隆号]-(多克隆)
[物种交叉反应]小鼠和大鼠(尚未测试其他物种)
[主持人]山羊
标签上指示的[浓度]
[配方] TBS

【Applications】
Immunohistochemistry( frozen section) 1 : 250-1,000
Immunohistochemistry( paraffin section) 1 : 250-1,000
Western Blot 1 : 1,000
Optimal concentration should be determined by each laboratory
for each application.
【Storage】
Store at -20℃ .  Avoid repeated freeze and thaw.
【Package】
100μL
【Recommended protocol( Immunohistochemistry frozen section)】
Wistar rat or ICR mouse was perfusion-fixed with 4% paraformaldehyde.
replaced sucrose, and prepared 25μm brain section
by microtome.
Wash : 0.3% TritonX-100 in PBS, 5 min × 3

Blocking : 1% BSA and 0.3% TritonX-100 in PBS, 2 hour, RT

Primary antibody : goat anti-Iba1 (1/1000), 1% BSA, and 0.3%

TritonX-100 in PBS, overnight, 4℃

Wash : 0.3% TritonX-100 in PBS, 5 min × 3

Secondary antibody : AlexaFluor488 anti-goat IgG (1/1000,
Jackson Immuno Research Laboratories #705-545-147), 1%
BSA, and 0.3% TritonX-100 in PBS, 2 hour, RT

Wash : 0.3% TritonX-100 in PBS, 5 min × 3

Mount
【Reference】 参考文献
1) Imai, Y., Ibata, I., Ito, D., Ohsawa, K. and Kohsaka, S. : Biochem.
Biophys. Res. Commun., 224, 855( 1996).

◆应用实例 1:免疫组织染色(荧光染色)

Immunohistochemistry (fluorescent staining)

03700482_img04.png

数据提供:国立长寿医疗研究中心榊原老师

样品:阿尔茨海默病模型小鼠(APPNL-G-F 小鼠)大脑新皮层冰冻切片

一抗:抗Iba1,山羊多克隆抗体(1:1,000)

二抗:Alexa Fluor488标记抗山羊IgG

Aβ染色:0.001 % FSB溶液(淀粉样蛋白染色荧光探针)

数据提供:创价大学理工学部中嶋老师

样本:大鼠(左)以及小鼠(右)大脑皮质冰冻切片

一抗:抗Iba1,山羊多克隆抗体(1:250)

二抗:Alexa Fluor488标记抗山羊IgG

◆应用实例 2:免疫组织染色(DAB染色)

03700482_omg02.png

样品:小鼠脑额叶石蜡切片

一抗:抗Iba1,山羊(1:1,000)

二抗:抗山羊IgG,生物素标记

抗原激活:10 mM柠檬酸盐缓冲液(pH 6),90°C,处理10 min

◆应用实例3:蛋白印迹 Western Blotting

03700482_omg03.png

数据提供:创价大学理工学部中嶋老师

样品:大鼠原代培养小胶质细胞  10 μg

样品:大鼠原代培养神经元 10 μg

样品:大鼠原代培养星形胶质细胞  10 μg

样品:大鼠大脑皮层  100 μg

一抗:抗Iba1,山羊(1:1,000)

二抗:抗山羊IgG,HRP标记

抗原 合成肽(Iba1 C端序列相同)
储存缓冲液 TBS
亚型 山羊IgG
物种交叉性 大鼠、小鼠
抗体浓度 0.5-0.6 mg/mL
应用 免疫组化(冰冻切片)1:250-1,000
免疫组化(石蜡切片)1:250-1,000
免疫印迹 1:1,000

◆产品信息

产品编号 产品名称 规格 包装
011-27991 Anti Iba1, Goat

抗Iba1,山羊源多克隆抗体

免疫化学用 100 μL

◆相关产品

产品编号 产品名称 规格 包装
019-19741 Anti Iba1, Rabbit (for Immunocytochemistry)
小胶质细胞/巨噬细胞特异性蛋白抗体(免疫组化)
免疫化学用 50 μg
013-27691 Anti Iba1, Rabbit(for Paraffin Section)
小胶质细胞/巨噬细胞特异性蛋白兔源抗体(石蜡切片)
免疫化学用 50 μg
016-26461 Anti Iba1, Rabbit, Biotin-conjugated
小胶质细胞/巨噬细胞特异性蛋白抗体(结合生物素)
免疫化学用 100 μL
013-26471 Anti Iba1, Rabbit, Red Fluorochrome(635)-conjugated
小胶质细胞/巨噬细胞特异性蛋白抗体(结合红色荧光素635)
免疫化学用 100 μL
016-20001 Anti Iba1,Rabbit (for Western Blotting)
小胶质细胞/巨噬细胞特异性蛋白抗体(免疫印迹)
免疫化学用 50 μg
012-26723 Anti Iba1, Monoclonal Antibody(NCNP24)
抗Iba1,单克隆抗体(NCNP24)(鼠源)
免疫化学用 10 μL
017-27591 Anti Human Iba1, Monoclonal Antibody(NCNP27)
抗人 Iba1,单抗(NCNP27)
免疫化学用 10 μL

Wako 和光纯药 011-27991 Anti Iba1, Goat Iba1抗体,山羊源多克隆抗体

Wako 和光纯药 011-27991 Anti Iba1, Goat  Iba1抗体,山羊源多克隆抗体

名称:Anti Iba1, Goat
用途:for Immunochemistry

Storage Condition : Keep at -20 degrees C.

Application Data

Immunohistochemistry (fluorescent staining)

03700482_img04.png

Immunohistochemistry of frozen section in mouse model of Alzheimer’s disease (APPNL-G-F mouse) brain cerebral neocortex using anti Iba1, goat polyclonal antibody at dilution of 1:1,000, and anti goat IgG, Alexa Flour 488-conjugated. Aβ were stained by 0.001% FSB solution(Amyloid fluorescent probe). The data were provided by Dr. Sakakibara, National Center for Geriatrics and Gerontology in Japan.

 

Immunohistochemistry of frozen section in rat(left) and mouse(right) brain cerebral cortex using anti Iba1, goat polyclonal antibody at dilution of 1:250, and anti goat IgG, Alexa Flour 488-conjugated.
The data were provided by Dr. Nakajima from Soka University in Japan.

Immunohistochemistry(DAB staining)

03700482_omg02.png

Sample: paraffin section of mouse brain frontal lobe
1st antibody: Anti Iba1, goat(1:1.000)
2nd antibody: Anti goat IgG, biotin-conjugated
Antigen retrieval: 10mM citric acid buffer(pH6.0), 90℃, 10min

Western Blotting

03700482_omg03.png

The data were provided by Dr. Nakajima from Soka University in Japan.

Sample:
Microglia from rat primary culture 10μg
Neuron from rat primary culture 10μg
Astrocyte from rat primary culture 10μg
Cerebral cortex from rat brain 100μg

1st antibody: Anti Iba1, goat(1:1.000)
2nd antibody: Anti goat IgG, HRP-conjugated

Antibody Profile

Antigen Synthetic peptide corresponding to C-terminal of Iba1
Buffer TBS
Species cross-reactivity Rat and Mouse
Antibody concentration 0.5-0.7 mg/mL
Application Immunohistochemistry(frozen section) 1:250-1,000
Immunohistochemistry(paraffin section) 1:250-1,000
Western blotting 1:1,000

Overview / Applications

Outline Iba1 is a calcium-binding protein with a molecular weight of 17,000 specifically expressed in macrophage and microglia.This product is goat polyclonal antibody that specifically recognize Iba1, and is available for a microglial marker.

Antigen: synthetic peptide(C-terminal of Iba1)
Cross-reactivity: mouse, rat
Applications:
Immunohistochemistry(frozen section) 1:250-1,000
Immunohistochemistry(paraffin section) 1:250-1,000
Western Blotting 1:1,000

Property

Appearance Liquid

Alias

  • Anti Iba1, Goat Polyclonal Antibody
    Anti AIF1, Goat Polyclonal Antibody
    Anti IRT1, Goat Polyclonal Antibody

 

 

WAKO 295-50201 DNA Extractor Kit 生物制药残余DNA抽提试剂盒

WAKO 295-50201 DNA Extractor Kit  生物制药残余DNA抽提试剂盒

DNA Extractor® Kit

for Genetic Research
Manufacturer :
FUJIFILM Wako Pure Chemical Corporation
Storage Condition :
Keep at 2-10 degrees C.
产品描述

血清、血浆可用。(更新日期:20190621)

传统的方式是采用蛋白酶-SDS的方法提取DNA,虽然能够获得高品质的DNA,但是后续需要用酚-氯仿进行处理,而酚-氯仿属于危险化学品,对人体有一定危害。

该试剂盒采用碘化钠法,能获得高质量和高回收率的DNA,抽提过程不需要酚-氯仿。

碘化钠代替酚-氯仿,溶解蛋白和脂类。糖原作为载体,异丙醇沉淀DNA。

只需将试剂加入离心管内即可,不像Qiagen的DNA抽提试剂盒采用柱吸附法。

产品回收率10pg左右。

文献报道的回收率是80%~90%。

Kit component

Kit component
50 tests
Sodium Iodide Solution 2 x 26 mL
Sodium N-Lauryl Sarcosinate Solution 1 x 1.2 mL
Washing Solution (A) 1 x 42 mL
Washing Solution (B) 2 x 40 mL
Glycogen Solution 1 x 0.1 mL

Features

  • Efficient recovery of trace DNA (100-1,000 fg)
  • Perform in a single tube for complete the process
  • Extraction completed in 60-90 minutes
  • Pretreatment protocol for samples containing high protein concentration
  • Adopts Sodium Iodide method*

*Sodium Iodide method is a residual DNA extraction technique that is described in The United States Pharmacopeia (USP) 42-NF37, <509> Residual DNA Testing.

Principle

Principle of DNA extraction

i. Cell membrane and cytoplasm are destroyed by surfactant, isolating cell nuclei.
ii. The nuclear membrane and nuclear protein are decomposed by protease to expose DNA.
iii. Protein and lipids are made soluble by the action of sodium iodide, and DNA is precipitated with isopropanol.

Extraction of viral DNA in serum and an infinitesimal amount of fungus-derived DNA in biological products

 

  • Extraction from serum

    01413_img01.png

    Operation time: 1 to 1.5 hours
    Sample amount: 100 μL/test

  • This kit is intended for use in extracting host cell-derived residual DNA remaining in biological components using the Sodium Iodide method. Extracted DNA can be quantified by qPCR. Use the DNA Extractor® kit for testing and management of the amount of DNA derived from host cells such as CHO cells, Escherichia coli, and yeast. DNA from any species extracted using this kit is suitable for using the Threshold Immunoassay Syste® provided by Molecular Devices, LLC. The kit can also be used for DNA extraction from serum.

Data

Recovery test of CHO cell-derived DNA

Purpose:
To examine the yields of CHO cell-derived DNA from culture supernatant.

Methods:
Using the kit, DNA was extracted from culture supernatants of PANC-1 cells to which 10 fg to 1 ng of CHO cell-derived DNA was added. A qPCR assay was then performed using the extracted DNA to obtain Cq values.
A qPCR was also performed using purified water to which CHO cell-derived DNA was added, without DNA extraction (standard conditions), to obtain Cq values. The DNA yield under each set of conditions was calculated based on a calibration curve generated from the results of the standard conditions.

Samples:
1. Purified water containing CHO cell-derived DNA (no DNA extraction): Standard conditions
2. DNA extracted from the culture supernatants of PANC-1 cells containing CHO cell-derived DNA, using the kit.

qPCR reagents:
・GeneAce SYBR® qPCR Mix α No ROX (Nippon Gene#319-07703)
・Optical Flat 8–Cap Strips for 0.2 mL tube strips/Plates (BIO-RAD#TCS0803)
・Hard-Shell® Thin-wall 96-well PCR plates (BIO-RAD#HSP9601)

Results:

1. Standard conditions 2. Culture supernatants
Added DNA (fg) Cq 1 Cq 2 Mean Cq 1 Cq 2 Mean
0 ND
10
100 36.89 ND 36.89 37.44 36.73 37.09
1,000 33.44 33.91 33.68 34.09 34.01
10,000 30.23 30.17 30.20 30.72 30.96 30.84
100,000 26.74 26.68 26.71 26.90 27.03 26.96
1,000,000 23.39 23.28 23.33 23.61 23.48 23.54

ND: Not detected

01413_img20.png

DNA added to culture supernatants (100 fg to 1 ng) was extracted with high yield.

This test was performed in accordance with Protocol #2 described in the manual.

Recovery test of E. coli-derived DNA

Purpose:
To examine the yields of E. coli-derived DNA.

Methods:
E. coli-derived DNA (10 fg to 1 ng) was added to purified water, and Cq values were measured before and after DNA extraction using the kit. Calibration curve was generated from mean Cq values of samples before the DNA extraction to calculate the yields after the extraction.

Samples:
Purified water containing E. coli-derived DNA

qPCR reagents:
・GeneAce SYBR® qPCR Mix α No ROX (Nippon Gene#319-07703)
・Optical Flat 8–Cap Strips for 0.2 mL tube strips/Plates (BIO-RAD#TCS0803)
・Hard-Shell® thin-wall 96-well PCR plates (BIO-RAD#HSP9601)

Results:

01413_img21.png

E. coli-derived DNA added to the samples (1 pg to 1 ng) was extracted with high yield.

This test was performed in accordance with Protocol #1 described in the manual.

Examples of using DNA Extractor® Kit

01413_img02.png

Reference

Ishizawa, M. et al., Nucleic Acids Res., 19, 5792 (1991).

Overview / Applications

Outline This is for research use only. Do not administer it to human. Molecular Biology-DNA Extraction Kits. Detects and quantitates contaminant DNA in serum and residual DNA in biopharmaceuticals. Employs a new extraction procedure for DNA purification from a single tube. This procedure, using Sodium Iodide (NaI) as a chaotropic agent, realizes DNA isolation of both high quality and high recovery from biological fluids without the use of phenol or chloroform. Complex and laborious manipulations are avoided when using this kit. In addition, this kit has a modified application for use with Molecular Devices’ Threshold(TM) System.
PRINCIPLE of DNA EXTRACTION: A high concentration of chaotropic reagent, NaI, and an anionic detergent participate in solubilization of the proteins and lipids contained in biological samples. After addition of isopropanol to the mixture, nucleic acids are co-precipitated with polysaccharide glycogen as a carrier, while other components remain soluble in the solution phase.

Ref.: Ishizawa, M., Kobayashi, T. and Matsuura, S., ”Simple procedure of DNA Isolation from Human Serum”, Nucleic Acids Res.19, 5792 (1991).

This method was listed on USP39-NF34, page 1413/Pharmacopeial Forum. Vol.41(4).

Purpose DNA extraction.

Related Information  其他相关产品:
Similar items list
DNA Extractor® Kit Series
Product List
Product Name
DNA Extractor® WB Kit (Sodium Iodide method) for Whole Blood DNA Extraction
DNA Extractor® SP kit for Genetic Research
DNA Extractor® TIS Kit for Genetic Research
DNA Extractor® FM Kit
8-OHdG Assay Preparation Reagent Set for Genetic Research

Wako 分散酶 Dispase CAS 9001-92-7

Wako 分散酶 Dispase CAS 9001-92-7

Dispase分散酶

Dispase分散酶是一类中性金属蛋白水解酶,可以回收生长在Matrigel基质上的细胞。与胰酶、胶原酶相比,它的作用更加温和,所以不会损伤细胞。此外,分散酶也可以用于组织分离。

来源:多粘芽孢杆菌表达的金属蛋白酶
使用指南:推荐浓度为10 U/cm2 BD Matrigel基质,如35 mm的培养皿推荐使用浓度为100U。
保存条件和效期:-20℃下冷冻保存,避免多次冻融。保质期详见产品包装。

Wako 分散酶 Dispase CAS 9001-92-7

383-02281 DISPASEⅡ 分散酶Ⅱ 1 g 9001-92-7
386-02271 DISPASE®Ⅰ 分散酶 10000 PU×6 9001-92-7

Roche公司的Dispase分散酶产品

货号 品名 规格
Roche-04942078001 Dispase® II (neutral protease, grade II) 1 g
Roche-04942078001 Dispase® II (neutral protease, grade II) 100mg

\

Wako和光纯药抑制剂 Inhibitor

Wako和光纯药抑制剂 Inhibitor

上海金畔生物代理Wako和光纯药全线产品,欢迎新老客户访问Wako和光纯药官网或者咨询我们获取更多相关产品信息。

Wako和光纯药代理

货号 品名 规格
307-50771 α-Amylase Inhibitor, From Wheat  α-淀粉酶抑制剂,来源于小麦 1MG
335-40623 Antipain 25MG
333-40624 Antipain 100MG
339-40621 Antipain 0.5MG
253-00471 YC-1 5MG
244-00721 Xestospongin C 100UG
210-00921 U-73343 5MG
213-00911 U-73122 5MG
211-01051 U0126 5MG
208-09223 Trypsin Inhibitor, from Soybean
胰蛋白酶抑制剂,大豆来源
1G
204-09225 Trypsin Inhibitor, from Soybean
胰蛋白酶抑制剂,大豆来源
5G
202-09226 Trypsin Inhibitor, from Soybean
胰蛋白酶抑制剂,大豆来源
500MG
202-09221 Trypsin Inhibitor, from Soybean
胰蛋白酶抑制剂,大豆来源
100MG
204-11991 Trichostatin A
曲古柳菌素A
5MG
200-11993 Trichostatin A
曲古柳菌素A
1MG
206-15471 Trichodion 100UG
209-14481 N-Tosyl-L-phenylalanine Chloromethyl Ketone(TPCK) 250MG
205-14483 N-Tosyl-L-phenylalanine Chloromethyl Ketone(TPCK) 1G
208-09181 (+/-)-alpha-Tocopherol Nicotinate
alpha-维生素E烟酸酯
5G
206-09182 (+/-)-alpha-Tocopherol Nicotinate
alpha-维生素E烟酸酯
25G
207-15641 TMP-153 500MG
164-17321 DL-threo-PPMP Hydrochloride
D,L-苏式-PPMP盐酸盐
25MG
161-17331 DL-threo-PPMP Hydrochloride
D,L-苏式-PPMP盐酸盐
50MG
206-10351 1-(2-Tetrahydrofuryl)-5-fluorouracil
1-(2-四氢呋喃基)-5-氟尿嘧啶
1G
202-10353 1-(2-Tetrahydrofuryl)-5-fluorouracil
1-(2-四氢呋喃基)-5-氟尿嘧啶
5G
209-12041 Tautomycin
泰托霉素
100UG
198-10281 Swainsonine
八倾吲嗪三醇
1MG
196-12703 Sulindac
舒林酸
50G
190-12701 Sulindac
舒林酸
10G
195-12491 Sulfuretin
硫黄菊素
20MG
333-31091 SUC-GLY-PRO-MCA 5MG
549-00286 STREPTOZOTOCIN 5G
543-00284 STREPTOZOTOCIN 1G
545-00283 STREPTOZOTOCIN 500MG
549-00281 STREPTOZOTOCIN 100MG
198-08515 Streptomycin Sulfate
硫酸链霉素
500G
192-08513 Streptomycin Sulfate
硫酸链霉素
100G
194-08512 Streptomycin Sulfate
硫酸链霉素
25G
196-08511 Streptomycin Sulfate
硫酸链霉素
5G
197-10251 Staurosporine
星形孢菌素
100UG
193-10253 Staurosporine
星形孢菌素
500UG
191-11533 Spectinomycin Dihydrochloride Pentahydrate
盐酸壮观霉素
1G
195-11531 Spectinomycin Dihydrochloride Pentahydrate
盐酸壮观霉素
5G
193-12051 Simvastatin
新伐他丁
25MG
194-13561 γ-Secretase Inhibitor X 1MG
188-01593 Roscovitine, 98.0+ % (HPLC)
细胞周期蛋白B激酶抑制剂
10MG
182-01591 Roscovitine, 98.0+ % (HPLC)
细胞周期蛋白B激酶抑制剂
1MG
181-01723 Resveratrol
白藜芦醇
500MG
185-01721 Resveratrol
白藜芦醇
100MG
183-01901 Radicicol
根赤壳菌素
1MG
165-20281 Protease Inhibitor Mixture-DMSO Solution for Fungal and Yeast Extracts 1ML
161-11493 Prednisolone
强的松龙
5G
165-11491 Prednisolone
强的松龙
1G
162-19821 Pravastatin Sodium Salt
普伐他汀
25MG
168-19823 Pravastatin Sodium Salt
普伐他汀
100MG
163-20341 PP-3
PP3酪氨酸激酶抑制剂
1MG
166-20331 PP 2, 94.0+ % (HPLC) 1MG
166-20294 Piroxicam
炎痛喜康
10G
162-20291 Piroxicam
炎痛喜康
1G
168-20293 Piroxicam
炎痛喜康
5G
160-17781 Phloretin, 98.0+ % (HPLC)
根皮素
250MG
160-11181 Pepsinostreptin
抑胃酶素
10MG
163-20101 PACOCF3
1,1,1-三氟-2-十七烷酮
10MG
144-07331 NS-398
COX-2选择性抑制剂NS-398
5MG
140-07333 NS-398
COX-2选择性抑制剂NS-398
25MG
147-07343 Niflumic Acid
尼氟酸
250G
141-07341 Niflumic Acid
尼氟酸
50G
145-06381 Nicardipine Hydrochloride, 99.0+ % (HPLC)
盐酸尼卡地平
1G
141-06383 Nicardipine Hydrochloride, 99.0+ % (HPLC)
盐酸尼卡地平
5G
145-06761 NF-κB inhibitor SN50 500UG
147-07201 (S)-(+)-Naproxen
(S)-(+)-萘普生
5G

Wako 074-06161 Glutelin, from Ric

Wako 074-06161 Glutelin, from Ric

Glutelin, from Rice(Yamadanishiki)
级别:for Cellbiology
代理商 : FUJIFILM Wako Pure Chemical Corporation
保存条件Storage Condition : Keep at 2-10 degrees C.

This product is for research use only. Do not administer it to human.

Glutelin is a major protein in rice, and defined as protein which dissolves in dilute acid and alkali.
In Japanese sake brewing, the byproducts of protein breakdown are considered to be a decisive factor in establishing flavor. The National Tax Agency of Japan has determined a special method to measure carboxypeptidase activity in koji mold (Aspergillus oryzae) used for fermentation. However, in recent years it has been reported that synthetic peptide substrates approved by this test are impractical in producing enzymatic activity from koji mold. To this extent, gluterin derived from rice has been shown to be an ideal solution to this problem.