Antibody Unit Protocol: ELISA

DAS ELISA Method Prepare coating antibody in coating buffer at recommended dilution and dispense (200 μL) into wells of an ELISA plate. Incubate 4hrs at 37oC. Wash plate x4 with wash buffer and blot dry. Prepare sample by homogenising plant material in extraction buffer 1/10 (w/v) and dispense (200 μL) into test wells. Incubate overnight at 4oC. Wash plate x4 with wash buffer and blot dry. Dilute antibody-AP conjugate at recommended dilution in extraction buffer and dispense (200 μL) into test wells. Incubate 2hrs 37oC. Wash plate x4 with wash buffer and blot dry. Prepare substrate 1mg/ml in substrate buffer and dispense (200 μL) into test wells. Incubate at ambient for 1-2hrs. Read OD of test wells at 405n. REAGENTS: Coating buffer, Wash buffer, Extraction buffer, Substrate buffer NB: The above protocol is used by us for the QC evaluations that are carried out by us on our reagents. Users have found that conditions and reagent volumes may be varied without substantially affecting the results obtained.