help with production of monoclonal antibodies

Kathryn Peters kathi at gwis.circ.gwu.edu
Mon Jul 11 16:06:10 EST 1994


In pursuit of my PhD, I have proposed, amongst other things, to raise
monoclonal antibodies against an extracellular epitope of a transmembrane
protein.  Although the other parts of this research are going well, the
production of these antibodies is not.  I have failed in several attempts
but I am determined to accomplish this aim.  I am ready to try the
procedure AGAIN.  Maybe  you could help!
What advice would you give me?  If you were to describe the factor(s) to
which you attribute your success with mabs, what would it (they) be?
In case you need to know:  Balb/c mice were injected with synthetic
peptide coupled to KLH; titers of the polyclonal serum were at least
1:1000 with ng quantities of peptide; SP2/O-AG14's were the fusion
partner; DMEM high glucose media was used and selection was with HAT; PEG
4000 was used for the fusion; I do get some hybridomas.
Would you send your response to my e-mail address?  Thank-you



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