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- Cell Signaling Technology
- 3999
- USA
- 2025年09月11日
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- 详细信息
- 文献和实验
- 技术资料
- 抗体英文名：
Streptavidin-HRP
- 库存：
大量
- 供应商：
CST
- 级别：
详见MSDS文件
- 保质期：
详见说明书
- 是否单克隆：
0
- 保存条件：
-20°c
- 规格：
1 ml/carrier free & custom formulation / quantity
| 规格： | 产品价格： | ￥请询价 | |
|---|---|---|---|
| 规格： | 1 ml | 产品价格： | ￥请询价 |
| 规格： | carrier free & custom formulation / quantity | 产品价格： | ￥请询价 |
Secondary Antibodies
Streptavidin-HRP #3999
Protocols
- 3999:
- Western Blotting
Specificity / Sensitivity
Streptavidin has a remarkably high affinity for its natural ligand, biotin. The complex and irregular structure of the biotin-binding site makes it highly optimized for biotin binding and confers great specificity to the streptavidin-biotin complexes (4).
Source / Purification
Streptavidin-HRP is purified by gel filtration. The conjugate product is free from unconjugated streptavidin and HRP.
Western Blotting
Western blot analysis of extracts from HeLa cells, untreated or treated with Paclitaxel #9807, using Phospho-Histone H3 (Ser10) (D2C8) Rabbit mAb (Biotinylated) #3642 and Streptavidin-HRP #3999 for detection.
Description
This Cell Signaling Technology product is useful for the detection of biotinylated proteins (1,2). Conjugation of horseradish peroxidase (HRP) to streptavidin is obtained by cross linking the amino groups on streptavidin with the carbohydrate groups on HRP.
Background
Streptavidin is a 53 kDa homotetramer isolated from Streptomyces avidinii for use in isolation and detection bioassays (3). Each streptavidin subunit forms high affinity non-covalent bonds with the vitamin biotin. Because of its strong non-covalent interaction with biotin, streptavidin can be used to detect and isolate biotinylated proteins (1,2).Chemiluminescent detection systems have emerged as the best all-around detection method for use with western blots and ELISA. These detection assays eliminate the hazards associated with radioactive materials and toxic chromogenic substrates. The speed and sensitivity of these methods are unequalled by traditional alternatives. Streptavidin-HRP is used with biotinylated proteins and specific chemiluminescent substrates to generate light signal. Streptavidin-HRP conjugates have a very high turnover rate, coupling high sensitivity with short reaction times.
- Updyke, T.V. and Nicolson, G.L. (1984) J Immunol Methods 73, 83-95.
- Buckie, J.W. and Cook, G.M. (1986) Anal Biochem 156, 463-72.
- Chaiet, L. and Wolf, F.J. (1964) Arch Biochem Biophys 106, 1-5.
- Reznik, G.O. et al. (1998) Proc Natl Acad Sci USA 95, 13525-30.
Application References
- Liang, J. et al. (2011) Mol Cell Biol 31, 2577-90. Applications: Western Blotting
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know !
Companion Products
- 9808 Phosphate Buffered Saline (PBS-20X)
- 9809 Phosphate Buffered Saline with Tween 20 (PBST-20X)
- 7004 TMB Substrate
- 7002 STOP Solution
- 9999 Nonfat Dry Milk
For Research Use Only. Not For Use In Diagnostic Procedures.
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文献和实验底物范围较广，包括二氨基联苯胺 (DAB)、氯萘酚和氨乙基咔唑等。 二氨基联苯胺/金属盐 DAB 是辣根过氧化物酶最敏感、常用的底物。它产生强烈的棕色产物，在水和醇中不溶解。多数情况下，推荐在 DAB 中加入不同的金属离子。当加入金属盐如钴、镍至底物液中，辣根过氧化物酶可以更加敏感。此类反应产物呈暗蓝灰色至黑色，且在水及醇中稳定。DAB/金属盐染色应用的组织染色范围较广。 1. 需要的溶液和特殊设备 DAB，0.05 mol/L Tris 缓冲液 (pH7.6), 0.3％ (W
问：有没有做辣根过氧化物标记的朋友啊，能不能把你们的步骤发给我。本人开始做，发现很多人用的浓度都不一样。答：1.1 试剂准备1) 10mg/mL HRP2) 0.06mol/L NaIO43) 0.16mol/L乙二醇4) 碳酸盐缓冲液(Na2CO3 0.7952g NaHCO3 1.4702g 溶于500mL蒸馏水中，调pH值到9.5)5) 2.5mg/mL NaBH46) 0.01mol/L磷酸盐缓冲液，pH7.41.2 操作步骤1. 10mg/mL取HRP VmL，溶液为棕黄色，加入
显色（HRP酶） 1．增强化学发光法(ECL) Ecl 显色原理：氨基苯二酰肼类主要是鲁米诺及异鲁米诺衍生物，是最常用的一类化学发光剂。鲁米诺（luminol，5＇-氨基-2，3-二氢-1，4-酞嗪二酮）的分子结构及化学反应式如下： 鲁米诺在免疫测定中既可用作标记物，也可用作过氧化物酶的底物。在Ecl底物中，含有H2O2和鲁米诺，在HRP（辣根过氧化物酶）的作用下，发出荧光来。 试验步骤： 1）将两种显色底物1：1等体积混合（一般各1ml/membrane）。
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