Principle of the Assay: Microtitration wells coated with murine anti-HIV-1 P24 capture antibody, are exposed to test specimens, which may contain p24 reactive determinants. After an incubation period, unbound components in the test sample are washed away. Specifically bound p24 reactive determinants react with a mouse anti-HIV-1 p24 conjugated with biotin during a second incubation period. Following a second wash cycle the biotinylated antibody is detected by the addition of a streptavidin HRP conjugate. Following a third wash cycle, specifically bound enzyme conjugate is detected by reaction with the Substrate solution, tetramethylbenzidine (TMB). The assay is measured spectrophotometrically to indicate the level of p24 reactive determinants present in a sample. Qualitative Analysis: Negative control should be 0.10 100pg/ml control should be 0.60 Sensitivity: 1.7pg/ml. Kit Components: H6005-10P1: Microtiter Strips, 1x96 wells Plastic microtitration wells coated with anti-HIV-1 p24 murine monoclonal antibody in foil pouch with desiccant. H6005-10P2: Positive Control 10ng/ml, 1x100ul H6005-10P3: Lysis Buffer, 1x6ml *H6005-10P4: Antibody (Biotin), 1x12ml H6005-10P5: Streptavidin (HRP), 1x12ml Containing 0.01% Bromonitrodioxane as preservative. H6005-10P6: Wash Buffer 20x, 2x60ml Tris buffered saline pH 7.8-8.0, containing 0.05% Tween 20. Must be diluted before use. H6005-10P7: Substrate (TMB) Solution, 1x12ml Ready to use. H6005-10P8: Stop Solution. 1N H2SO4, 1x12ml Storage and Stability: Aliquot and store *H6005-10P4 at -20C. Store other components at 4C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
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