2 replies to this topic

[kroger]

hi,guys:
    I am doing human cancer cell Elisa now, and I try to check if my protein will interact with the receptors on cell surface.I  use HRPgoat anti-mouse IgG conjugate as the secondary antiboday. I am just  confused: Is it necessary to use the blocking buffer to block the unspecific binding for the secondary antibody?
   thanks a lot

Edited by kroger, 28 January 2005 - 11:06 AM.

[blossom]

Without knowing the specific details of your ELISA.  I only block before adding my standards or samples to the wells.  The sequence is 1) capture antibody, 2) blocking, 3) samples, 4) detecting antibody, 5)developing agent.

Hope this answered your question!

[kroger]

blossom, on Jan 28 2005, 02:15 PM, said:

Without knowing the specific details of your ELISA.  I only block before adding my standards or samples to the wells.  The sequence is 1) capture antibody, 2) blocking, 3) samples, 4) detecting antibody, 5)developing agent.

Hope this answered your question!

thank you very much, it's my first time to try Elisa,I am thinking if I don't use blocking buffer,Will the unspecific binding have great effect to my result?