CD40/TNFRSF5 Antibody (82111) [Unconjugated]

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.

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

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.

• Reactivity: Hu
• Applications: Flow, CyTOF-ready, Func, ICC/IF
• Clone: 82111
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

CD40/TNFRSF5 Antibody (82111) [Unconjugated] Summary

• 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.
• Source: N/A
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Mouse
• Gene: CD40
• Purity Statement: Protein A or G purified from hybridoma culture supernatant
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • Agonist Activity
  • CyTOF-ready
  • Flow Cytometry 2.5 ug/10^6 cells
  • Immunocytochemistry 8-25 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for CD40/TNFRSF5 Antibody (82111) [Unconjugated]

• 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

Background

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).

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for CD40/TNFRSF5 Antibody (MAB6321)(9)

Publications using MAB6321

• I Harder, M Münchhalfe, G Andrieux, M Boerries, B Grimbacher, H Eibel, ME Maccari, S Ehl, J Wienands, J Jellusova, K Warnatz, B Keller Dysregulated PI3K Signaling in B Cells of CVID Patients Cells, 2022-01-28;11(3):. 2022-01-28 [PMID: 35159274] (Stimulation, Human)
• Chae Yoon Lim, Jae Hyuck Chang, Won Sun Lee, Jeana Kim, Il Young Park CD40 Agonists Alter the Pancreatic Cancer Microenvironment by Shifting the Macrophage Phenotype toward M1 and Suppress Human Pancreatic Cancer in Organotypic Slice Cultures Gut and Liver 7/15/2022 [PMID: 34933280]
• G Brummer, W Fang, C Smart, B Zinda, N Alissa, C Berkland, D Miller, N Cheng CCR2 signaling in breast carcinoma cells promotes tumor growth and invasion by promoting CCL2 and suppressing CD154 effects on the angiogenic and immune microenvironments Oncogene, 2019-12-11;0(0):. 2019-12-11 [PMID: 31827233] (Flow Cytometry, Mouse)
• PM Schlievert, MP Cahill, BS Hostager, AJ Brosnahan, AJ Klingelhut, FA Gourronc, GA Bishop, DYM Leung Staphylococcal Superantigens Stimulate Epithelial Cells through CD40 To Produce Chemokines MBio, 2019-03-19;10(2):. 3/19/2019 [PMID: 30890614]
• Hatterer E, Barba L, Noraz N et al. Co-engaging CD47 and CD19 with a bispecific antibody abrogates B-cell receptor/CD19 association leading to impaired B-cell proliferation. MAbs 12/19/2018 [PMID: 30569825]
• Maddur M, Sharma M, Hegde P, Stephen-Victor E, Pulendran B, Kaveri S, Bayry J Human B cells induce dendritic cell maturation and favour Th2 polarization by inducing OX-40 ligand. Nat Commun, 2014-06-09;5(0):4092. 2014-06-09 [PMID: 24910129] (Neutralization, Human)
• Heine G, Drozdenko G, Grun J, Chang H, Radbruch A, Worm M Autocrine IL-10 promotes human B-cell differentiation into IgM- or IgG-secreting plasmablasts. Eur J Immunol, 2014-03-11;44(6):1615-21. 2014-03-11 [PMID: 24643722] (Neutralization, Human)
• 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 Deficient EBV-specific B- and T-cell response in patients with chronic fatigue syndrome. PLoS ONE, 2014-01-15;9(1):e85387. 2014-01-15 [PMID: 24454857] (Functional Assay, Human)
• 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 Fibrosis in human adipose tissue: composition, distribution, and link with lipid metabolism and fat mass loss. Diabetes, 2010-08-16;59(11):2817-25. 2010-08-16 [PMID: 20713683] (IHC-P, Human)

FAQs for CD40/TNFRSF5 Antibody (MAB6321). (Showing 1 - 2 of 2 FAQs).

1. I wanna know does CD40 antibody for IHC-P comes with positive control slide?
   • Our CD40 antibodies do not come with positive control slides however you can use any type of primary carcinoma cell as a positive control. I recommend breast carcinoma as this is what we have tested our CD40 antibodies on.
2. We are studying CD40L in our system. It has been known that both CD40 and CD11b are receptors for CD40L. We need CD40 antibody to neutralize CD40 binding to CD40L, then we may prove what is function for CD11b binding to CD40L. Do you have any CD40 Ab which has block function only between CD40 and CD40L, but not between CD11b and CD40L?
   • Unfortunately, none of our CD40 antibodies have been specifically tested for this application.

Bioinformatics

• Gene Symbol: CD40
• Uniprot:
  • P25942()