Experiments can not be explained. mAb binding site problem!!! - 未名空间MITBBS历史存档

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Experiments can not be explained. mAb binding site problem!!!

Experiments can not be explained. mAb binding site problem!!!# Biology - 生物学

s*y2011-04-05 07:04

1 楼

purchased a mAb against protein with domains of
A-B-C-D-E
The mAb is produced by recombinant protein of B plus a small region on A.
Now, I purified the GST-A, GST-B, GST-C, GST-D, GST-E, and GST-Full
length.
When testing binding by ELISA, only GST-Full length and GST-B can be
recognized by this antibody.
But, in SDS-PAGE and wester blot, GST-A, GST-B, GST-C, GST-D and GST-Full
length, but not GST and GST-E, can be recognized by this mAb.
Totally lost for an applicable explanation, please help. thanks.

e*s2011-04-05 07:04

2 楼

How about sequence similarity between these domains?
Elisa, all domains are in soluble conformation. A's small region could
be folded inside.
In WB, all domains and proteins are denatured. If there are some highly
conserved region between B,C, and D, then I think you got an explanation.

A.
Full

【在 s*********y 的大作中提到】

: purchased a mAb against protein with domains of
: A-B-C-D-E
: The mAb is produced by recombinant protein of B plus a small region on A.
: Now, I purified the GST-A, GST-B, GST-C, GST-D, GST-E, and GST-Full
: length.
: When testing binding by ELISA, only GST-Full length and GST-B can be
: recognized by this antibody.
: But, in SDS-PAGE and wester blot, GST-A, GST-B, GST-C, GST-D and GST-Full
: length, but not GST and GST-E, can be recognized by this mAb.
: Totally lost for an applicable explanation, please help. thanks.