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Antibody Cross reactivity
phoenix red
Friday, 30 May 2008 07:33 UTC
have inherited some primary antibody in my lab which seems to cross react giving me tremendous background on my westerns. So…the recombinant protein was harvested from e-coli which was poorly purified resulting in e-coli protein being present in the product. this was shot into a bunny and thus the Ab was produced. Now that i use it probe a archeal protein expression it lights up all over the western. My guess is that its probably groel or Groes. However i need to clean up the antibody ASAP so as to get rid of the bacterial stuff!!!. does any one know of any protocols or papers with protocols that can help me. I am in a real fix here and would appreciate it if you could help me out in any way !
thank you
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Replies
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We have a protocol in the peer-reviewed section of our content that might be of use to you:
Purification of antibodies using the synthetic affinity ligand absorbent MAbsorbent A2P
From the abstract it might be specific to antibodies from ovine serum, but I cannot think at the moment why it could not be adapted to others.
Parts of the protocol that you don’t need a site-licence to access:
Table 1. MAbsorbent A2P—product specifications
Table 2. Overview of purification steps
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