CD40 protein

CD40-muIg -R-PE

Cat. Num. AAA666791

• Reactivity: Human
• Applications: Flow Cytometry, Functional Assay, ELISA
• Synonyms: CD40; CD40-muIg -R-PE; Human CD40 muIg/R-PE Fusion Protein; CD40 protein

• Host: CHO cells
• Reactivity: Human
• Form/Format: 50 mM Sodium Phosphate pH 7.5, 500 mM Potassium Chloride, 150mM NaCl, 15% Glycerol, 0.2% BSA.
• Concentration: Volume: 0.2 ml (varies by lot)
• Sequence Length: 117

• Applicable Applications for CD40 protein: Flow Cytometry (FC/FACS), ELISA (EIA)
• Application Notes: 1:20 (or use 4 ul of concentrated stock per 5 x 105-cell test)
• 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: Ficoll prepared human peripheral blood mononuclear cells were stimulated 6 hours in culture in the presence of 10 ng/ml PMA and 1uM Ionomicin to upregulate CD154 expression. Five x 105 cells per tube were washed and incubated 45 minutes on ice with 80 ul of a 1:20 dilution (10 ug/ml) of CD40-muIg/R-PE. They were then washed three times, fixed and analyzed by FACS. Cells stained positive with a mean shift of 1.4 log10 fluorescent units when compared to a Mouse IgG2a/R-PE negative control at a similar concentration. Binding was blocked when cells were pre incubated 10 minutes with 20 ul of 0.5 mg/ml anti-CD154 antibody.
• Conjugation: PE
• Production: Protein A purified recombinant protein from tissue culture supernatant was conjugated to R-Phycoerythrin through a sulfo-ester linkage. Unconjugated protein was removed using size exclusion chromatography. Protein conjugate is at a concentration of 0.2 mg/ml with an A565/A280 ratio of 3.7.
• Buffer: 50 mM Sodium Phosphate pH 7.5, 500 mM Potassium Chloride, 150mM NaCl, 15% Glycerol, 0.2% BSA, 0.04% NaN3 (as a preservative).
• Preparation and Storage: Store at 2 - 5 degree C. Do not freeze! Protect from light.; 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). CD40 muIg fusion protein is recognized by anti-human CD40 antibody (catalog #300-020) and binds to human CD154. This binding can be blocked by anti-CD154 antibody (catalog #353-020).

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.

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; positive regulation of isotype switching to IgG isotypes; platelet activation; 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; positive regulation of B cell proliferation; protein kinase B signaling cascade; protein complex assembly; positive regulation of transcription from RNA polymerase II promoter; positive regulation of protein amino acid phosphorylation; inflammatory response; defense response to virus; regulation of immunoglobulin secretion

Disease: Immunodeficiency With Hyper-igm, Type 3