发信人: royluo ( 罗马情结), 信区: Biology
标 题: Re: 一张NITROCELLULOSE膜可以做多少次strip?
发信站: The unknown SPACE (Wed May 14 01:52:04 2003), 站内信件


blot stripping buffer:
100mM beta-Mecaptoethanol
2% SDS
625mM Tris.Hcl pH6.8

To make 50 ml stripping buffer:

41.5ml ddH2O
5ml 20% SDS
3.125ml 1M Tris.HCl pH6.8
0.352ml beta-Mecaptoethanol

To use the buffer to strip the blot:

1. incubate the blot in the stripping buffer for
30 min at 70 celcius degree
2. Wash the blot with PBST five times

Note: this protocol's condition is very harsh. It is very
possible that not only the antibody but also some of the protein
immoblized on the blot will be stripped away. Milder conditions
like room temperature incubation and lower concentration of SDS
can be tried.


【 在 windysea (神仙眷侣~我怀念你爱上我的第一天) 的大作中提到: 】
: co-admire,
: can you give us the exact recipe and conditions (such as temp, time, w/ or w/o
: shaking)?
: 【 在 mystery (Polaris) 的大作中提到: 】
: : admire


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※ 来源:．The unknown SPACE bbs.mit.edu．[FROM: 129.49.]