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Protein detection problem under reducing condition

Protein detection problem under reducing condition - Protein Forum

Protein detection problem under reducing condition - Protein Forum


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  #1  
Old 06-27-2003, 07:40 AM
Mauro Bongrazio
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Default Protein detection problem under reducing condition



" reducing.." and "..non-reducing conditions " refer to the target protein and not
to the antibody. The Ab was used for immunoblot detection of target protein in cell
lysates resolved by SDS-PAGE under reducing (no signal) or non-reducing (signal)
conditions.
Mauro

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Old 07-23-2003, 04:57 PM
Michael Schaefer
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Default Protein detection problem under reducing condition

> to the antibody. The Ab was used for immunoblot detection of target protein in cell

Did you immunize the animal with a peptide containing more than the terminal cysteine
which may be required for hooking it up to KLH?

If this is the case, some of the binding AB-species may recognize the oxidized form of
the cysteines. If your sample has a similar structure including disulfide bonds, these
more avidly binding antibodies may be predominant.

Michael

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