Bovine Serum Albumin ELISA Assay


The Bovine Serum Albumin ELISA Assay is used as an analytical tool for the quantitative determination of Bovine Serum Albumin in serum, plasma and other biological samples. 

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
Reactivity: Bovine
Detection Method: Colorimetric
Storage: 2-8 °C

Assay Principle

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. 

Related Products

Bovine IgG1 ELISA Assay
Human Albumin ELISA Assay
Bovine Haptoglobin ELISA Assay

Additional Information

Assay Background

The Bovine Serum Albumin ELISA Assay is used for assessing the specific biomarker in samples analytes which may be serum, plasma and cell culture supernatant as validated with the kit. The kit employs a competitive ELISA technique. 

Assay Procedure

1. It is strongly recommended that all Standards and Samples be run in duplicates or triplicates. A standard curve is required for each assay. 

2. Add 50 ul prepared Standards and Samples to respective wells. 

3. Pipette 50 ul Biotinylated Bovine Serum Albumin Antigen Working Solution to all wells. 

4. Cover the plate with a sealer and incubate for 60 minutes at 37°C. 

5. Aspirate and wash plate 4 times with diluted Wash Buffer (1X) and blot residual buffer by firmly tapping plate upside down on absorbent paper. Wipe of any liquid from the bottom outside of the microtiter wells as any residue can interfere in the reading step. 

6. Pipette 100 ul Streptavidin:HRP Conjugate Working Solution to all wells. Mix well. 

7. Cover the plate with a sealer and incubate for 30 minutes at 37°C. 

8. Aspirate and wash as per Step (5) above. 

9. Pipette 100 ul TMB Substrate in all the wells. 

10. Incubate the plate at 37°C for 10 minutes. DO NOT SHAKE or else it may result in higher backgrounds and worse precision. 

11. Pipette 100 ul of Stop Solution to all wells. The wells should turn from blue to yellow in color. 

12. Read the absorbance at 450 nm with a microplate within 10-15 minutes after addition of Stop solution. 

Package Inserts

Please note: All documents above are for reference use only and should not be used in place of the documents included with this physical product. If digital copies are needed, please contact us.

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