...Using the Indirect ELISA Method Antibody Validation Using the Indirect ELISA Method You may also like... ...of each well at 450 nm on the spectrophotometer. Use the spectrophotometer to generate a standard curve... we list the specific equipment, reagents, and methods that we use in our lab at Addgene. Equipment and...an automated ELISA plate washer. Equipment Spectrophotometer compatible with 96-well plates 1–10 µL single...PBS, 1X pH 7.4, VWR 45000-446 TMB (3,3',5,5'-tetramethyl-[1,1'-biphenyl]-4,4'-diamine) substrate kit, ...