BISC 220 Lab 8 expected results

BISC 220 Lab 8 expected results - Chapter 8 ELISA technique...

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Chapter 8: ELISA technique Specifications Coating Buffer : 0.1% Sodium Bicarbonate, pH 8.6. Coated antigen : Salmonella typhimurium Washing buffer : 0.01M Phosphate Buffered Saline pH 7.4 with Tween 20 (.05%) Blocking buffer : 0.01M Phosphate Buffered Saline pH 7.4 with 1% Bovine serum albumin (BSA) Primary antibody : Abcam ab35156 rabbit polyclonal IgG (100x dilution in blocking buffer) Enzyme-labeled secondary antibody : Sigma A3687 polyclonal goat anti-rabbit IgG, alkaline phosphatase (AP) conjugated (1000x dilution in blocking buffer) Substrate : BCIP/NBT (alkaline phosphatase; chromogenic – color change) BCIP : 5-bromo-4-chloro-3-indolyl phosphate NBT : p-nitroblue tetrazolium chloride. Results Just as the lab manual describes, well 12 (your control) should be regarded as colorless (though color can evolve slowly) because there was absolutely no patient serum added. Since the “patient serum” contains antibody, well 1-11 would show blue color gradually changing from dark to light, to colorless.
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