Human TNFβ ELISA Kit

Catalog No.: BEK1215

Size: 96T

Range: 62.5 pg/ml-4000 pg/ml

Sensitivity < 5 pg/ml

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

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

Introduction

Tumor necrosis factor-beta (TNF-beta), previously called lymphotoxin-alpha(LTA), is a member of the tumor necrosis factor family , is a cytokine produced by lymphocytes. It is a glycoprotein with a relative molecular mass (Mr) of 60,000-70,000. LTA is highly inducible, secreted, and exists as homotrimeric molecule. It forms heterotrimers with lymphotoxin-beta which anchors lymphotoxin-alpha to the cell surface. LTA mediates a large variety of inflammatory, immunostimulatory, and antiviral responses. It is also involved in the formation of secondary lymphoid organs during development and plays a role in apoptosis.

Principle of the Assay

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-P- Cadherin polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-TNFβ 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 TNFβ amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of TNFβ can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-Human TNFβ antibody
2. Lyophilized human TNFβ standards: 2 tubes (10 ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-human TNFβ 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