Mouse CD40/ TNFRSF5 ELISA Kit

Catalog No.: BEK1029

Size: 96T

Range: 15.6 pg/ml-1000 pg/ml

Sensitivity < 1 pg/ml

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

Application: For quantitative detection of CD40 in mouse serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

CD40 is a costimulatory protein found on antigen presenting cells and is required for their activation. It is expressed on the surface of all mature B cells. This protein receptor is a member of the TNF-receptor superfamily. It has been found to be essential in mediating a broad variety of immune and inflammatory responses including T cell-dependent immunoglobulin class switching, memory B cell development, and germinal center formation. The interaction of this receptor and its ligand is found to be necessary for amyloid-beta-induced microglial activation, and thus is thought to be an early event in Alzheimer disease pathogenesis.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-CD40 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-CD40 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 CD40 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of CD40 can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-Mouse CD40 antibody
2. Lyophilized Mouse CD40 standards: 2 tubes (10 ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-Mouse CD40 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