Bovine Serum Albumin ELISA Assay
The Bovine Serum Albumin ELISA Assay is For Research Use Only
Size: 8×12 wells
Sensitivity: 161.2 ng/ml
Calibrator Range: 0 ng/ml – 30000 ng/ml
Sample Type: Cell Culture Supernatant and Other Biological Preparations
Detection Method: Colorimetric
Storage: 2-8 °C
The method employs competitive enzyme-linked immunosorbent assay (ELISA) technique to assay the level of Bovine Serum Albumin in samples. Standards or Samples competes with the biotinylated Bovine Serum Albumin, to form a complex with the Bovine Serum Albumin antibody coated microtiter well. Wells are washed to remove the excess conjugate and Streptavidin:HRP Conjugate is added to the microplate and incubated. After incubation and a washing step TMB Substrate, are added. Blue color develops on incubation and the reaction is stopped with a Stop Solution to form a yellow color. The concentration of the Bovine Serum Albumin in the samples is inversely proportional to the yellow color developed (absorbance) in the wells.
Sample Preparation & Storage
Specimens should be clear and non-hemolyzed. Samples should be run at a number of dilutions to ensure accurate quantitation.
1. Extract as soon as possible after specimen collection as per relevant procedure. The samples should be tested as soon as possible after the extraction. Alternately the extracted samples can be kept in -20°C. Avoid repeated freeze-thaw cycles.
2. Serum- Coagulate at room temperature for 10-20 minutes; centrifuge for 20-min at 2000-3000 rpm. Remove the supernatant. If precipitation appears, recentrifuge.
3. Plasma- Use EDTA or citrate plasma as an anticoagulant, mix for 10-20 minutes; centrifuge for 15-min at 2000-3000 rpm. Remove the supernatant carefully. If precipitation appears, recentrifuge.
4. Cell Culture Supernatant- Collect sample in a sterile container. Centrifuge for 20-mins at 2000-3000 rpm. Remove the supernatant carefully. When examining the components within the cell, dilute cell suspension with PBS (pH 7.2-7.4), if cell concentration is greater than 1 million/ml. Damage the cells by repeated freeze-thaw cycles to release intracellular components. Centrifuge for 20-min at 2000-3000 rpm. If precipitation appears, centrifuge again.
5. Tissue Samples- Rinse tissues in PBS (pH 7.4) to remove excess blood thoroughly and weigh before homogenization. Mince tissues and homogenize them in PBS (pH7.4) with a glass homogenizer on ice. Thaw at 2-8°C or freeze at -20°C. Centrifuge at 2000-3000 RPM for approximately 20 minutes and collect the supernatant carefully.
Note: Grossly hemolyzed samples are not suitable for use in this assay.
Bovine IgG1 ELISA Assay
Human Albumin ELISA Assay
Bovine Haptoglobin ELISA Assay