CD40 protein

CD40-muIg -Biotin

Cat. Num. AAA666814

• Reactivity: Human
• Applications: Flow Cytometry, Functional Assay, ELISA
• Synonyms: CD40; CD40-muIg -Biotin; Human CD40 muIg/Biotin; CD40 protein

• Host: CHO cells
• Reactivity: Human
• Form/Format: 50 mM Sodium Phosphate pH 7.5, 100 mM Potassium Chloride, 150mM NaCl, 5% Glycerol, 0.2% BSA.
• Concentration: 0.5 mg/ml (varies by lot)
• Sequence Length: 117

• Applicable Applications for CD40 protein: Flow Cytometry (FC/FACS), ELISA (EIA)
• Molecular Structure: A soluble fusion protein consisting of the extracellular (193aa) domain of human CD40 fused to murine IgG2a Fc (233aa).
• Transfectant Cell Line: CHO
• Performance: Jurkat-4G cells were stimulated 6 hours in the presence of PMA and Ionomycin (to increase CD154 expression). Five x 105 cells per tube were washed and incubated 45 minutes on ice with 80 mul of CD40-muIg/Biotin at a concentration of 5 mug/ml. Cells were washed twice and incubated with 2 degree reagent Streptavidin/R-PE, after which they were washed three times, fixed and analyzed by FACS. Cells stained positive with a mean shift of 0.78 log10 fluorescent units when compared to a buffer control at a similar concentration. Binding was blocked when cells were pre blocked 10 minutes with 20 mul of anti-CD154 antibody at a concentration of 0.5 mg/ml.
• Conjugation: Biotin
• Production: Fusion protein from (low FBS containing) tissue culture supernatant of transfectants was purified using affinity and size exclusion chromatography), and reacted with NHS-Biotin. Unconjugated Biotin was removed from conjugate by diafiltration.
• Buffer: 50 mM Sodium Phosphate pH 7.5, 100 mM Potassium Chloride, 150mM NaCl, 5% Glycerol, 0.2% BSA, 0.04% NaN3 (as a preservative).
• Preparation and Storage: Store at 2 - 5 degree C. Freeze/Thawing is not recommended.; Product should retain activity for at least 12 months after shipping date when stored as recommended.

Testing Data

Related Product Information for CD40 protein

Information: Human CD40 is a member of the tumor necrosis factor (TNF) receptor family and is present on all B cells except plasma cells. CD40 is also found on some epithelial cells, carcinomas and lymphoid dendritic cells. CD40 plays an important role in B cell activation and the interaction with its ligand CD154 (CD40 Ligand) is essential for isotype switching (1). The CD40-CD154 interaction appears to have a role in artherosclerosis and cardiovascular disorders (3,4,5). CD40-muIg fusion protein binds to cell surface CD154 in Flow cytometry.

References

1) T.A. Foy, et al, (1996) Annu Rev Immunol 14: 591-617. 2) M.E. Ozaki, et al, (1997) J Immunol 159: 214-221. 3) F. Mach, et al, (1997) Proc Natl Acad Sci USA 94:1931-1936. 4) U. Schonbeck, et al, (2000) PNAS USA 97: 7458-7463. 5) U. Schonbeck, et al, (2001) Circulation 104: 2266-2268.

NCBI and Uniprot Product Information

• NCBI GI #: 114053977
• Molecular Weight: 13,158 Da
• NCBI Official Full Name: CD40, partial
• UniProt Protein Name: CD40
• Protein Family: CD40 ligand
• UniProt Entry Name: Q09LL4_HUMAN

NCBI Description

This gene is a member of the TNF-receptor superfamily. The encoded protein is a receptor on antigen-presenting cells of the immune system and is essential for mediating a broad variety of immune and inflammatory responses including T cell-dependent immunoglobulin class switching, memory B cell development, and germinal center formation. AT-hook transcription factor AKNA is reported to coordinately regulate the expression of this receptor and its ligand, which may be important for homotypic cell interactions. Adaptor protein TNFR2 interacts with this receptor and serves as a mediator of the signal transduction. 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. Mutations affecting this gene are the cause of autosomal recessive hyper-IgM immunodeficiency type 3 (HIGM3). Multiple alternatively spliced transcript variants of this gene encoding distinct isoforms have been reported. [provided by RefSeq, Nov 2014]

Uniprot Description

CD40: a member of the TNF-receptor superfamily. This receptor for CD40L mediates a broad variety of immune and inflammatory responses including T cell-dependent immunoglobulin class switching, memory B cell development, and germinal center formation. Defects in CD40 are the cause of hyper-IgM immunodeficiency type 3 (HIGM3). HIGM3 is an autosomal recessive disorder which includes an inability of B cells to undergo isotype switching, one of the final differentiation steps in the humoral immune system, an inability to mount an antibody-specific immune response, and a lack of germinal center formation. Two alternatively spliced isoforms have been reported. Isoform I is a type I membrane protein; isoform II is secreted.

Protein type: Membrane protein, integral; Receptor, cytokine

Chromosomal Location of Human Ortholog: 20q12-q13.2

Cellular Component: extracellular space; cell surface; intracellular membrane-bound organelle; integral to plasma membrane; cytoplasm; plasma membrane; external side of plasma membrane

Molecular Function: signal transducer activity; protein binding; enzyme binding; ubiquitin protein ligase binding; receptor activity; antigen binding

Biological Process: B cell proliferation; platelet activation; positive regulation of isotype switching to IgG isotypes; regulation of immune response; positive regulation of I-kappaB kinase/NF-kappaB cascade; positive regulation of interleukin-12 production; immune response-regulating cell surface receptor signaling pathway; activation of NF-kappaB transcription factor; cellular calcium ion homeostasis; positive regulation of MAP kinase activity; positive regulation of tyrosine phosphorylation of Stat1 protein; protein kinase B signaling cascade; positive regulation of B cell proliferation; positive regulation of transcription from RNA polymerase II promoter; protein complex assembly; positive regulation of protein amino acid phosphorylation; inflammatory response; defense response to virus; regulation of immunoglobulin secretion

Disease: Immunodeficiency With Hyper-igm, Type 3