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标题: HMGB1 antibody 高迁移移蛋白1（ab79823）的热修复是柠檬酸还是EDTA？ [打印本页]

作者: promotion 时间: 2011-10-13 22:30 标题: HMGB1 antibody 高迁移移蛋白1（ab79823）的热修复是柠檬酸还是EDTA？

HMGB1 antibody 高迁移移蛋白1（ab79823）的热修复是柠檬酸还是EDTA？如果是EDTA的话，PH是多少？浓缩液稀释的比例大约是多少？一抗的修复时间多少？请提供对于做该一抗的相关要求，谢谢！

作者: shanghainese 时间: 2011-10-14 11:46

引用:

原帖由 promotion 于 2011-10-13 22:30 发表
HMGB1 antibody 高迁移移蛋白1（ab79823）的热修复是柠檬酸还是EDTA？如果是EDTA的话，PH是多少？浓缩液稀释的比例大约是多少？一抗的修复时间多少？请提供对于做该一抗的相关要求，谢谢！

非常简单，问abcom。

作者: promotion 时间: 2011-10-14 13:45

谢谢

作者: shanghainese 时间: 2011-10-16 11:47

HMGB1 Immunohistochemical staining

Four micrometers of tissue sections were de-paraffinized, rehydrated, and treated with an antigen retrieval solution (10 mmol/L sodium citrate buffer, pH 6.0). The sections were incubated with a dilution of 1:500 rabbit anti-HMGB1 antibody (Abcam, UK) overnight at 4°C, and then incubated with 1:1000 dilution of biotinylated secondary antibody, followed by avidin-biotin peroxidase complex (DAKO, Carpinteria, CA) according to the manufacturer's instructions. Finally, tissue sections were incubated with 3', 3'-diaminobenzidine (Sigma-Aldrich, Co., Ltd) until a brown color developed, and then the sections were counterstained with Harris' modified hematoxylin. In negative controls, primary antibodies were omitted. Hepatocytes that have brown staining in nucleus area represent normal location of HMGB1, while hepatocytes with brown staining in both cytoplasmic and nucleus area were defined as cytoplasmic translocation of HMGB1. At least 10 high-power fields were chosen randomly, and >1000 cells were counted for each section. Values represent percentages of hepatocytes with HMGB1 cytoplasmic translocation in all hepatocytes counted in each group.

http://www.biomedcentral.com/1471-230X/11/21

作者: promotion 时间: 2011-10-17 11:45 标题: 回复 4# 的帖子

万分感谢斑竹这么用心帮助。

作者: 天涯无风 时间: 2011-11-17 20:30

用柠檬酸行的，用EDTA肯定行

作者: shanghainese 时间: 2011-11-18 22:55

引用:

原帖由 天涯无风 于 2011-11-17 20:30 发表
用柠檬酸行的，用EDTA肯定行

不一定。

作者: 天涯无风 时间: 2011-11-21 12:16

引用:

原帖由 shanghainese 于 2011-11-18 22:55 发表

不一定。

举个例子

作者: shanghainese 时间: 2011-11-21 13:50

希望各位在做免疫组化时按照说明书上的步骤作，特别是抗原修复。

作者: shanghainese 时间: 2011-11-21 13:56

引用:

原帖由 天涯无风 于 2011-11-21 12:16 发表

举个例子

请您说明一下，有多少单克隆抗体用柠檬酸行的，用EDTA肯定行的，并列出他们的clone name。

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