I might have wrongly used HRP-conjugated antibody instead of non-conjugated one. But if so, I think protein G would not bind this antibody anymore and there should be no band at all. Am I right?
Thanks.
Edited by gyma, 28 June 2010 - 02:05 AM.
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#1
gyma
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Posted 28 June 2010 - 02:03 AM
I did IP and got no band but heavy and light chains, and samples include a group of positive control. does that mean the antibody didnt work?
I might have wrongly used HRP-conjugated antibody instead of non-conjugated one. But if so, I think protein G would not bind this antibody anymore and there should be no band at all. Am I right?
Thanks.
I might have wrongly used HRP-conjugated antibody instead of non-conjugated one. But if so, I think protein G would not bind this antibody anymore and there should be no band at all. Am I right?
Thanks.
#2
Inmost sun
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Posted 02 July 2010 - 11:22 AM
[quote name='gyma' date='Jun 28 2010, 10:03 AM' post='77297']
I did IP and got no band but heavy and light chains, and samples include a group of positive control. does that mean the antibody didnt work?
I might have wrongly used HRP-conjugated antibody instead of non-conjugated one. But if so, I think protein G would not bind this antibody anymore and there should be no band at all. Am I right?
Thanks.
[/quote
Full binding of protein G may be hindered by HRP for steric reason; additionally, unspecific binding of proteins to HRP may increase unspecific precipitations
I did IP and got no band but heavy and light chains, and samples include a group of positive control. does that mean the antibody didnt work?
I might have wrongly used HRP-conjugated antibody instead of non-conjugated one. But if so, I think protein G would not bind this antibody anymore and there should be no band at all. Am I right?
Thanks.
[/quote
Full binding of protein G may be hindered by HRP for steric reason; additionally, unspecific binding of proteins to HRP may increase unspecific precipitations
