Help to increase the protein recovery of BSA
Ang Ching Seng
93462036 at comet.np.ac.sg
Fri Dec 1 01:15:35 EST 1995
I'm a final year student doing my last year project which is to raise
antibody in mice against the pesticide Methemidophos(Met). The approach is to
reduce BSA to expose its thiol group so that the Met molecule which is being
linked to a cross linker (Sulfo-SMCC which is from pierce) is able to cross
link the BSA (BSA acts as a carrier molecule).
I used 90umol of Dithiolthreiol(DTT) to reduce the BSA followed by
reccipitation using 5% TCA ( I've tried higher concentration of TCA) after
washing with 2mM EDTA in dH20 the final resuspension is in 0.05M phosphate
buffer 8M Urea 2mM EDTA. My recovery rate is always at the 40-50% range and
i wonder if anyone has any alternative methods and i would be very grateful
if you could E-mail me.
I would be very much appreciate if anyone could tell me any other ways of
raising the antibodies against the pesticide THANK YOU !
From Ang Ching Seng
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