BSA cross reaction

[Bernard Murray]

In article <95284.102007U12201 at uicvm.uic.edu>, U12201 at uicvm.uic.edu says...
>
>In article <Pine.SOL.3.91.951009190429.21464A-100000 at suma3.reading.ac.uk>, 
Supannee Chitpinityol <skrchitl at reading.ac.uk> says:
>>
>>Hi!
>>
>>I have made polyclonal antichymosin but there is a cross reaction with
>>BSA, possibly the contamination from the original protein used for
>>injection into the rabbit. Could anyone  give me an idea how to get rid
>>this contaminant antibody. I heard that someone using BSA agarose to bind
>>the antiBSA. Have anyone experience using this method.
>>
>>Many thanks in advance.
>>
>>Supannee.
>
>One way is to immobilize some BSA on a column, then pass the
>antibody solution over it.
>Keld Sorensen

Just wondering.  Have you eliminated the possibility that it is your *second*
antibody that is cross-reacting with BSA? (please don't be insulted if I am
stating the obvious)  I have used some (including that supplied with
Amersham's ECL kit) that show this kind of effect.  I refrain from using
milk/blotto etc. and use BSA for blocking and in the incubation and washing
buffer and this helps considerably.  If it is the primary antibody then I
agree with the poster above and suggest back-absorbing it with immobilised
"unwanted" antigen.
		Good luck!
			Bernard
Bernard Murray, Ph.D.
bernard at elsie.nci.nih.gov  (National Cancer Institute, NIH, Bethesda MD, USA)