Human BAFF/TNFSF13B ELISA Kit

Catalog No.: BEK1009

Size: 96T

Range: 62.5 pg/ml - 4000 pg/ml

Sensitivity < 2 pg/ml

Storage and Expiration: Store at 2-8℃ for 6 months, or at -20℃ for 12 months. 

Application: For quantitative detection of BAFF in serum, plasma, body fluids, tissue lysates or cell culture supernates. 

Introduction

B-cell activating factor (BAFF) also known as tumor necrosis factor ligand superfamily member 13B is a cytokine that belongs to the tumor necrosis factor (TNF) ligand family. It was expressed in B cell lineage cells, and acts as a potent B cell activator. It has been also shown to play an important role in the proliferation and differentiation of B cells.As an immunostimulant, BAFF is necessary for maintaining normal immunity.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-BAFF polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-BAFF polyclonal antibody was used as detection antibodies. The standards, test samples and biotin conjugated detection antibody were added to the wells subsequently, and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color

product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the BAFF amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of BAFF can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-Human BAFF antibody
2. Lyophilized BAFF standards: 2 tubes (10ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-Human BAFF antibody (Concentrated): 130μl. Dilution: 1:100
5. Antibody diluent buffer: 12ml
6. Avidin-Biotin-Peroxidase Complex (ABC) (Concentrated): 130μl. Dilution: 1:100
7. ABC diluent buffer: 12ml
8. TMB substrate: 10ml
9. Stop solution: 10ml
10. Wash buffer (25X): 30ml

Note: Reconstitute standards and test samples with Kit Component 3.

Material Required But Not Provided

1. 37℃ incubator
2. Microplate reader (wavelength: 450nm)
3. Precise pipette and disposable pipette tips
4. Automated plate washer
5. ELISA shaker
6. 1.5ml of Eppendorf tubes
7. Plate cover
8. Absorbent filter papers
9. Plastic or glass container with volume of above 1L