This reagent is intended for research use only and is designed to quantify bovine serum albumin (BSA) in bovine serum, plasma, and related liquid samples. The assay is based on the double-antibody sandwich ELISA method. A microplate is pre-coated with purified BSA-specific antibodies, forming a solid-phase antibody. The sample is then added, allowing BSA to bind to the immobilized antibodies. Subsequently, horseradish peroxidase (HRP)-labeled BSA is introduced, forming an immune complex of antibody-antigen-enzyme-labeled antibody. After thorough washing, the substrate TMB is added. Under HRP catalysis, TMB turns blue and then changes to yellow upon acid addition. The intensity of the color is directly proportional to the BSA concentration in the sample. Absorbance is measured at 450 nm using a microplate reader, and the BSA concentration is determined by comparing the OD values to a standard curve.
The kit includes components such as the enzyme-labeled plate, standards, diluents, substrates, and wash solutions, all stored at 2–8°C. Proper sample preparation is essential for accurate results. Serum samples should be centrifuged after clotting, while plasma requires anticoagulants like EDTA or sodium citrate. Urine and body fluids must be centrifuged before testing. Cell culture supernatants should be collected and centrifuged, while tissue samples need homogenization in PBS. All samples should be processed promptly, and if not tested immediately, they should be stored at -20°C without repeated freezing and thawing.
The procedure involves serial dilution of standards, sample addition, incubation, washing, enzyme labeling, color development, and termination. Each step must be carefully followed to ensure accuracy. The standard curve should be generated each time, and samples with high concentrations should be diluted appropriately. The sealing film is for single-use only to prevent cross-contamination, and all waste materials should be handled as biohazardous. The kit performance ensures a linear regression coefficient R ≥ 0.99 and intra- and inter-assay variability below 9% and 11%, respectively. The detection range is 3–70 μg/L. The kit has a shelf life of 6 months when stored at 2–8°C. Always refer to the English manual for the most accurate instructions.
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