Human CD40/TNFRSF5 Antibody

Catalog # Availability Size / Price Qty
MAB6321-SP
MAB6321-100
MAB6321-500
CD40/TNFRSF5 in Human PBMCs.
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Product Details
Citations (9)
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Supplemental Products
Reviews (1)

Human CD40/TNFRSF5 Antibody Summary

Species Reactivity
Human
Specificity
Detects human CD40/TNFRSF5 in direct ELISAs and Western blots. In direct ELISAs, does not cross-react with recombinant human (rh) 4‑1BB, rhCD27, rhCD30, recombinant mouse CD40, rhDR3, rhDR6, rhEDAR, rhFas, rhGITR, rhHVEM, rhLTR beta, rhNGF R, rhOPG, rhRANK, rhTAJ, rhTNF RI, or rhTNF RII.
Source
Monoclonal Mouse IgG2B Clone # 82111
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
Mouse myeloma cell line NS0-derived recombinant human CD40/TNFRSF5
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
Flow Cytometry
2.5 µg/106 cells
Human whole blood CD19+ B cells
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 0.035-0.175 μg/mL.
 
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 
Immunocytochemistry
8-25 µg/mL
See below

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

Immunocytochemistry CD40/TNFRSF5 antibody in Human PBMCs by Immunocytochemistry (ICC). View Larger

CD40/TNFRSF5 in Human PBMCs. CD40/TNFRSF5 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Mouse Anti-Human CD40/TNFRSF5 Monoclonal Antibody (Catalog # MAB6321) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces and cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

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. Mouse Anti-Human CD40/TNFRSF5 Monoclonal Antibody (Catalog # MAB6321) stimulates human B cell proliferation in the presence of Recombinant Human IL-4 (Catalog # 204-IL) in a dose-dependent manner, as measured by Resazurin (Catalog # AR002). The ED50 for this effect is typically 0.035-0.175 µg/mL

Flow Cytometry View Larger

Detection of CD40/TNFRSF5 in PBMC lymphocytes by Flow Cytometry PBMC lymphocytes were stained with Mouse Anti-Human CD19 APC‑conjugated Monoclonal Antibody (Catalog # FAB4867A) and either (A) Mouse Anti-Human CD40/TNFRSF5 Monoclonal Antibody (Catalog # MAB6321) or (B) isotype control antibody (Catalog # MAB0041) followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). View our protocol for Staining Membrane-associated Proteins.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 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.

9 Citations: Showing 1 - 9
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  1. CD40 Agonists Alter the Pancreatic Cancer Microenvironment by Shifting the Macrophage Phenotype toward M1 and Suppress Human Pancreatic Cancer in Organotypic Slice Cultures
    Authors: Chae Yoon Lim, Jae Hyuck Chang, Won Sun Lee, Jeana Kim, Il Young Park
    Gut and Liver
  2. Staphylococcal Superantigens Stimulate Epithelial Cells through CD40 To Produce Chemokines
    Authors: PM Schlievert, MP Cahill, BS Hostager, AJ Brosnahan, AJ Klingelhut, FA Gourronc, GA Bishop, DYM Leung
    MBio, 2019-03-19;10(2):.
  3. Dysregulated PI3K Signaling in B Cells of CVID Patients
    Authors: I Harder, M Münchhalfe, G Andrieux, M Boerries, B Grimbacher, H Eibel, ME Maccari, S Ehl, J Wienands, J Jellusova, K Warnatz, B Keller
    Cells, 2022-01-28;11(3):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Stimulation
  4. CCR2 signaling in breast carcinoma cells promotes tumor growth and invasion by promoting CCL2 and suppressing CD154 effects on the angiogenic and immune microenvironments
    Authors: G Brummer, W Fang, C Smart, B Zinda, N Alissa, C Berkland, D Miller, N Cheng
    Oncogene, 2019-12-11;0(0):.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  5. Human B cells induce dendritic cell maturation and favour Th2 polarization by inducing OX-40 ligand.
    Authors: Maddur M, Sharma M, Hegde P, Stephen-Victor E, Pulendran B, Kaveri S, Bayry J
    Nat Commun, 2014-06-09;5(0):4092.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  6. Autocrine IL-10 promotes human B-cell differentiation into IgM- or IgG-secreting plasmablasts.
    Authors: Heine G, Drozdenko G, Grun J, Chang H, Radbruch A, Worm M
    Eur J Immunol, 2014-03-11;44(6):1615-21.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  7. Deficient EBV-specific B- and T-cell response in patients with chronic fatigue syndrome.
    Authors: Loebel M, Strohschein K, Giannini C, Koelsch U, Bauer S, Doebis C, Thomas S, Unterwalder N, von Baehr V, Reinke P, Knops M, Hanitsch L, Meisel C, Volk H, Scheibenbogen C
    PLoS ONE, 2014-01-15;9(1):e85387.
    Species: Human
    Sample Types: Whole Cells
    Applications: Functional Assay
  8. Fibrosis in human adipose tissue: composition, distribution, and link with lipid metabolism and fat mass loss.
    Authors: Divoux A, Tordjman J, Lacasa D, Veyrie N, Hugol D, Aissat A, Basdevant A, Guerre-Millo M, Poitou C, Zucker JD, Bedossa P, Clement K
    Diabetes, 2010-08-16;59(11):2817-25.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-P
  9. Co-engaging CD47 and CD19 with a bispecific antibody abrogates B-cell receptor/CD19 association leading to impaired B-cell proliferation.
    Authors: Hatterer E, Barba L, Noraz N et al.
    MAbs

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Human CD40/TNFRSF5 Antibody
By Anonymous on 06/02/2019
Application: WB Sample Tested: Cancer cell lysates,Min6 mouse insulinoma whole cell lysate Species: Human