Human CD40/TNFRSF5 Antibody

Name: Human CD40/TNFRSF5 Antibody
Brand: R&D Systems
SKU: AF632

R&D Systems, Inc. a Bio-Techne Brand

Catalog # AF632

Citations (5)

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Key Product Details

Species Reactivity:	Human
Immunogen:	Mouse myeloma cell line NS0-derived recombinant human CD40 Glu21-Arg193 Accession # P25942
Applications:	Functional Assay, Simple Western, Western Blot
Label:	Unconjugated
Source:	Polyclonal Goat IgG
Specificity:	Detects human CD40 in direct ELISAs and Western blots.

View Full Technical Details for Human CD40/TNFRSF5 Antibody

Applications for Human CD40/TNFRSF5 Antibody

	Recommended Concentration	Sample
Western Blot	2 µg/mL	See below
Simple Western	50 µg/mL	Raji human Burkitt's lymphoma cell line and Daudi human Burkitt's lymphoma cell line
Agonist Activity	Measured in a cell proliferation assay using B cell enriched human peripheral blood lymphocytes in the presence of IL-4. Banchereau, J. et al. (1991) Science 251:70. The ED₅₀ for this effect is typically 3-12 ng/mL.

Please Note: Optimal dilutions should be determined by each laboratory for each application.

Scientific Data Examples for Human CD40/TNFRSF5 Antibody

	Western Blot Detection of Human CD40/TNFRSF5 by Western Blot. Western blot shows lysates of Raji human Burkitt's lymphoma cell line and Daudi human Burkitt's lymphoma cell line. PVDF membrane was probed with 2 µg/mL of Goat Anti-Human CD40/TNFRSF5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF632) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). A specific band was detected for CD40/TNFRSF5 at approximately 40-45 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
	Agonist Activity Human CD40/TNFRSF5 Antibody Stimulates Cell Proliferation in Human B Cells. Goat Anti-Human CD40/TNFRSF5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF632) stimulates human B cell proliferation in the presence of Recombinant Human IL-4 (204-IL) in a dose-dependent manner, as measured by Resazurin (AR002). The ED₅₀ for this effect is typically 3-12 ng/mL
	Simple Western Detection of Human CD40/TNFRSF5 by Simple Western^TM. Simple Western lane view shows lysates of Raji human Burkitt's lymphoma cell line and Daudi human Burkitt's lymphoma cell line, loaded at 0.2 mg/mL. A specific band was detected for CD40/TNFRSF5 at approximately 57 kDa (as indicated) using 50 µg/mL of Goat Anti-Human CD40/TNFRSF5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF632). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Formulation, Preparation and Storage

Purification	Antigen Affinity-purified
Endotoxin level	<0.10 EU per 1 μg of the antibody by the LAL method.
Reconstitution	Reconstitute at 0.2 mg/mL in sterile PBS. Reconstitution Buffer Available: Size / Price Qty Loading... RB01:
Formulation	Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Shipping	The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage	Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 12 months from date of receipt, -20 to -70 °C as supplied. 1 month, 2 to 8 °C under sterile conditions after reconstitution. 6 months, -20 to -70 °C under sterile conditions after reconstitution.

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Background: CD40/TNFRSF5

CD40 is a type I transmembrane glycoprotein belonging to the TNF receptor superfamily. The mature hCD40 consists of a 172 amino acid (aa) extracellular domain, a 22 aa transmembrane region and a 62 aa cytoplasmic domain (1). Human and mouse CD40 share 62% aa identity. CD40 is expressed in B cells, follicular dendritic cells, dendritic cells, activated monocytes, macrophages, endothelial cells, vascular smooth muscle cells, and several tumor cell lines (2). The extracellular domain has the cysteine-rich repeat regions, which are characteristic for many of the receptors of the TNF superfamily. Interaction of CD40 with its ligand, CD40L, leads to aggregation of CD40 molecules, which in turn interact with cytoplasmic components to initiate signaling pathways. Early studies on the CD40-CD40L system revealed its role in humoral immunity. Interaction between CD40L on T cells and CD40 on B cells stimulated B cell proliferation and provided the signal for immunoglobulin isotype switching (3). Mutations in the CD40L gene, which resulted in a CD40L molecule unable to interact with CD40, are responsible for the hyper-IgM syndrome (4). Cross-linking of CD40 with antibodies or by CD40 binding to CD40L produces cell type-specific responses which include costimulation and induction of proliferation, induction of cytokine production, rescue from apoptosis, and upregulation of adhesion molecules (5). Some of the early events of intracellular signaling by the CD40-CD40L system include the association of the CD40 with TRAFs and the activation of various kinases (6 - 8).

Item	Value
References	Torres, R.M. and E.A. Clark (1992) J. Immunol. 148:620. Schonbeck, U. et al. (1997) J. Biol. Chem. 272:19569. Armitage, R.J. et al. (1993) J. Immunol. 150:3671. Callard, R.E. et al. (1993) Immunol. Today 14:559. Stout, R.D. and J. Suttles (1996) Immunol. Today 17:487. Pullen, S.S. et al. (1999) Biochemistry 38:10168. Faris, M. et al. (1994) J. Exp. Med. 179:1923. Hanissian, S.H. and R.S Geha (1997) Immunity 6:379.
Entrez Gene IDs	958 (Human); 21939 (Mouse); 171369 (Rat); 102118696 (Cynomolgus Monkey)
Alternate Names	B cell surface antigen CD40, B-cell surface antigen CD40, Bp50B cell-associated molecule, CD40, CD40 antigen, CD40 molecule, TNF receptor superfamily member 5, CD40 type II isoform, CD40L receptor, CDw40, MGC9013, TNFRSF5, TNFRSF5CD40 antigen (TNF receptor superfamily member 5), nerve growth factor receptor-related B-lymphocyte activation molecule, p50, tumor necrosis factor receptor superfamily member 5, tumor necrosis factor receptor superfamily, member 5

Citations for Human CD40/TNFRSF5 Antibody (5)

Showing 1 - 5 of 5 citations Showing All

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Citations are publications that use Bio-Techne products. Selected citations for Human CD40/TNFRSF5 Antibody include:

Species: Human
Sample Types: Whole Tissue
Applications: IHC

T Schmidt et al. (2020), Children with oligoarticular juvenile idiopathic arthritis have skewed synovial monocyte polarization pattern with functional impairment-a distinct inflammatory pattern for oligoarticular juvenile arthritis Arthritis Res. Ther., 22(1):186.
PMID: 32787920
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot

Zhan Y et al. (2016), Expression, purification and biological characterization of the extracellular domain of CD40 from Pichia pastoris. BMC Biotechnol, 16(1):8.
PMID: 26809818
Species: Human
Sample Types: Whole Cells
Applications: Neutralization

Karnell J et al. (2014), CD19 and CD32b differentially regulate human B cell responsiveness. J Immunol, 192(4):1480-90.
PMID: 24442430
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot

Dranitzki-Elhalel M et al. (2007), CD40.FasL inhibits human T cells: evidence for an auto-inhibitory loop-back mechanism. Int. Immunol., 19(4):355-63.
PMID: 17314083
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot

Nemoto E et al. (2002), Disruption of CD40/CD40 ligand interaction with cleavage of CD40 on human gingival fibroblasts by human leukocyte elastase resulting in down-regulation of chemokine production. J. Leukoc. Biol., 72(3):538-45.
PMID: 12223522

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Human CD40/TNFRSF5 Antibody

R&D Systems, Inc. a Bio-Techne Brand

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Applications for Human CD40/TNFRSF5 Antibody

Scientific Data Examples for Human CD40/TNFRSF5 Antibody

Western Blot

Agonist Activity

Simple Western

Formulation, Preparation and Storage

Reconstitution Calculator

Background: CD40/TNFRSF5

Citations for Human CD40/TNFRSF5 Antibody (5)

Product Documents for Human CD40/TNFRSF5 Antibody

Product Datasheets

COA

Certificate of Analysis

SDS

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