Re: testing antibody dilution
From your descriptions of your antibody titers I
would think your 1:5000 dilution may be too strong.
The 1:7000 dilution may be the optimal dilution and
you may wish to try a 1:6000 since you saw the nuclear
staining drop a bit in intensity in the positive
control. Be certain you are blocking your sections
appropriately before applying your antibody to
discourage non-specific staining. Though the
non-specific cytoplasmic staining you are describing
may be caused by endogenous biotin. I would suggest
trying non Biotinylated secondary system or a polymer
kit. Zymed offers a Non-Biotin Amplification detection
kit, and a PicTure Plus polymer detection system. Both
would be an excellent choice to combat such a problem.
Good luck and best wishes,
James Syrewicz II
South Central Representative
Zymed Laboratories, Inc.
Voice mail: 800-874-4494 ext. 8629
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