dilution of antibody when used with super g blocking buffer

2013-12-01

azim58 - dilution of antibody when used with super g blocking buffer

question for Grace Biolabs

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Hi Grace Biolabs representative, I am curious about what I should dilute
my antibody in when I am using it after the Super G Blocking Buffer has
been used. I have nitrocellulose slides spotted with tumor cDNA
expression library cell lysate which will be blocked with Super G. I will
then probe this slide with antibody. The typical incubation buffer that
we use in our lab consists of 5 mL 30% BSA, 25 uL of Tween 20, 5 mL 10X
PBS, and 40 mL H20. However, I wonder if it would be best for me to avoid
incubating the antibody with BSA since the blocking buffer had no BSA in
it and Super G Blocking Buffer was used. In this situation, perhaps I
should just dilute the antibody in PBS for best results? Or maybe PBS
with some Tween 20? Thanks for any feedback you have to offer!

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Grace Biolabs reply

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Hi Kurt,
I recently was forwarded your question regarding incubation buffer for
primary antibody following Super G blocking of nitrocellulose. Normally,
here for in house assays I have used PBS with 0.1% Tween20. Using BSA
should not be a problem either though, and for longer incubations, may
help to prevent protein aggregation. I would test both to see what
performs best for your application but I do not foresee any issues with
either. Please let me know if you have any further questions and I would
be happy to answer them.
Best Regards,
Michael

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