Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Flow Cytometry, Immunocytochemistry, Agonist Activity, CyTOF-ready

Cited:

Immunohistochemistry-Paraffin, Neutralization, Flow Cytometry, Bioassay, Functional Assay, Stimulation

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 82111
View all CD40/TNFRSF5 Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human CD40/TNFRSF5
Glu21-Arg193
Accession # P25942

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.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Human CD40/TNFRSF5 Antibody

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

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.
Human CD40/TNFRSF5 Antibody Stimulates Cell Proliferation antibody in Human B Cells.

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

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.

Applications for Human CD40/TNFRSF5 Antibody

Application
Recommended Usage

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.

Flow Cytometry

2.5 µg/106 cells
Sample: Human whole blood CD19+ B cells

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed human peripheral blood mononuclear cells (PBMCs)

Reviewed Applications

Read 1 review rated 4 using MAB6321 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

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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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.

Alternate Names

CD40, TNFRSF5

Entrez Gene IDs

958 (Human); 21939 (Mouse); 171369 (Rat); 102118696 (Cynomolgus Monkey)

Gene Symbol

CD40

UniProt

P25942

Additional CD40/TNFRSF5 Products

Product Documents for Human CD40/TNFRSF5 Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human CD40/TNFRSF5 Antibody

For research use only

Citations for Human CD40/TNFRSF5 Antibody

Customer Reviews for Human CD40/TNFRSF5 Antibody (1)

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  • Human CD40/TNFRSF5 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Min6 mouse insulinoma whole cell lysate and Cancer cell lysates
    Species: Human
    Verified Customer | Posted 06/02/2019
    Human CD40/TNFRSF5 Antibody MAB6321

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