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Human CD40/TNFRSF5 Antibody

Catalog #: AF632
5 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
AF632
AF632-SP
Detection of Human CD40/TNFRSF5 by Western Blot.
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Product Details
Citations (5)
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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human CD40/TNFRSF5 Antibody Summary

Species Reactivity
Human
Specificity
Detects human CD40 in direct ELISAs and Western blots.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
Mouse myeloma cell line NS0-derived recombinant human CD40
Glu21-Arg193
Accession # P25942
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.
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.
Label
Unconjugated

Applications

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. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Human CD40/TNFRSF5 antibody by Western Blot. View Larger

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 antibody in Human B Cells. View Larger

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 View Larger

Detection of Human CD40/TNFRSF5 by Simple WesternTM. 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.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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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.

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).

References
  1. Torres, R.M. and E.A. Clark (1992) J. Immunol. 148:620.
  2. Schonbeck, U. et al. (1997) J. Biol. Chem. 272:19569.
  3. Armitage, R.J. et al. (1993) J. Immunol. 150:3671.
  4. Callard, R.E. et al. (1993) Immunol. Today 14:559.
  5. Stout, R.D. and J. Suttles (1996) Immunol. Today 17:487.
  6. Pullen, S.S. et al. (1999) Biochemistry 38:10168.
  7. Faris, M. et al. (1994) J. Exp. Med. 179:1923.
  8. 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 antigen; CD40 molecule, TNF receptor superfamily member 5; CD40 type II isoform; CD40; CD40L receptor; CDw40; MGC9013; nerve growth factor receptor-related B-lymphocyte activation molecule; p50; TNFRSF5; TNFRSF5CD40 antigen (TNF receptor superfamily member 5); tumor necrosis factor receptor superfamily member 5; tumor necrosis factor receptor superfamily, member 5

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Citations for Human CD40/TNFRSF5 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

5 Citations: Showing 1 - 5
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  1. Children with oligoarticular juvenile idiopathic arthritis have skewed synovial monocyte polarization pattern with functional impairment-a distinct inflammatory pattern for oligoarticular juvenile arthritis
    Authors: T Schmidt, E Berthold, S Arve-Butle, B Gullstrand, A Mossberg, F Kahn, AA Bengtsson, B Månsson, R Kahn
    Arthritis Res. Ther., 2020-08-12;22(1):186.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  2. Expression, purification and biological characterization of the extracellular domain of CD40 from Pichia pastoris.
    Authors: Zhan Y, Wei Y, Chen P, Zhang H, Liu D, Zhang J, Liu R, Chen R, Zhang J, Mo W, Zhang X
    BMC Biotechnol, 2016-01-25;16(1):8.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  3. CD19 and CD32b differentially regulate human B cell responsiveness.
    Authors: Karnell J, Dimasi N, Karnell F, Fleming R, Kuta E, Wilson M, Wu H, Gao C, Herbst R, Ettinger R
    J Immunol, 2014-01-17;192(4):1480-90.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  4. CD40.FasL inhibits human T cells: evidence for an auto-inhibitory loop-back mechanism.
    Authors: Dranitzki-Elhalel M, Huang JH, Sasson M, Rachmilewitz J, Parnas M, Tykocinski ML
    Int. Immunol., 2007-02-20;19(4):355-63.
    Species: Human
    Sample Types: Cell Lysates, Whole Cells
    Applications: Western Blot
  5. 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.
    Authors: Nemoto E, Tada H, Shimauchi H
    J. Leukoc. Biol., 2002-09-01;72(3):538-45.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot

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