Determine secondary antibodies that will be needed and prepare Carrier and blocking solutions with sera from the host of the secondary. (ie. Goat anti Rabbit secondary antibody-- make Carrier and block solutions with normal goat serum)
A. | Carrier Solution: Makes 1L To 800ml of dH2O, add 100ml 10X PBS, pH 7.2, add: |
10g BSA 15ml 20% Triton (0.3% Final) 10ml Normal Serum (To match host of secondary antibody, 0.1% final) Bring to 1000ml final volume with dH2O. Check pH and adjust as necessary to 7.4. |
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B. | Blocking Solution: 20% Normal serum (To match host of secondary antibody) |
20ml Normal Serum |
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