Overview

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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.

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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.

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Range: 15.6 pg/ml-1000 pg/ml

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Sensitivity < 1 pg/ml

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Storage and Expiration: Store at 2-8℃ for 4 months, or at -20℃ for 8 months.

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

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Kit components

1.     One 96-well plate pre-coated with anti-Human CD40 antibody
2.     Lyophilized Human CD40 standards: 2 tubes (10ng / tube)                    
3.     Sample / Standard diluent buffer: 30ml
4.     Biotin conjugated anti-Human CD40 antibody (Concentrated): 130 μl. Dilution: 1:100
5.     Antibody diluent buffer: 12 ml
6.     Avidin-Biotin-Peroxidase Complex (ABC) (Concentrated): 130 μl. Dilution: 1:100
7.     ABC diluent buffer: 12 ml
8.     TMB substrate: 10 ml
9.     Stop solution: 10 ml
10.   Wash buffer(25X): 30 ml

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

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Gene Information

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OMIM: 109535
Swiss Port: P25942
Gene ID: 958

 

Other Information

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Please check datasheet for protocol.