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Key Product Details

Species Reactivity

Validated:

Human

Applications

Flow Cytometry

Label

Alexa Fluor 405 (Excitation = 405 nm, Emission = 421 nm)

Antibody Source

Monoclonal Mouse IgG2B Clone # 40804

Product Summary for Human CD40 Ligand/TNFSF5 Alexa Fluor® 405-conjugated Antibody

Immunogen

E. coli-derived recombinant human CD40 Ligand/TNFSF5
Leu50-Leu261
Accession # P29965

Specificity

Detects human CD40 Ligand/TNFSF5 in direct ELISAs and Western blots. In direct ELISAs and Western blots, no cross-reactivity with recombinant human (rh) CD27 Ligand, rhCD30 Ligand, or recombinant mouse CD40 Ligand is observed.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Applications for Human CD40 Ligand/TNFSF5 Alexa Fluor® 405-conjugated Antibody

Application
Recommended Usage

Flow Cytometry

0.25-1 µg/106 cells
Sample: Human peripheral blood mononuclear cells treated with PMA and Ca2+ ionomycin
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Formulation

Supplied 0.2 mg/mL in a saline solution containing BSA and Sodium Azide.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: CD40 Ligand/TNFSF5

CD40 Ligand (CD40L), now renamed TNFSF5 but also known as CD154, TRAP and gp39, is a 34‑39 kDa type II transmembrane glycoprotein that belongs to the TNF superfamily (1‑3). Human CD40L is 261 amino acids (aa) in length and consists of a 22 aa cytoplasmic domain, a 24 aa transmembrane segment, and a 215 aa extracellular region that consists of multiple beta-strands and one N‑linked glycosylation site (4, 5). Although carbohydrates are present, they are not necessary for activity (6). As with other TNF superfamily members, CD40L will exist as a trimer, both as a membrane bound and soluble form (6‑8). The soluble form is 18 kDa in size and about 150 aa in length, and arises from intracellular proteolytic processing. As a trimer, the soluble form is bioactive (7‑9). Multiple mutations and alternate splice forms of CD40L exist, often associated with pathology and leading to truncated or nontrimerizable forms of CD40L (8). While CD40L is a ligand for CD40, the ligation of CD40L by CD40 initiates bidirectional signaling in both CD40 and CD40L expressing cells (10). The extracellular region of human CD40L is 99%, 88%, 86%, 82%, 75% and 75% aa identical to the extracellular regions of CD40L in rhesus monkey, bovine, porcine, canine, mouse and rat, respectively. CD40L binds to both CD40 and to integrin  alphaIIb beta3 (CD41) (3, 11). In the cell membrane, it also associates with a unique splice variant of CD28 (CD28i) that may facilitate CD40L signal transduction (12). CD40L is expressed by monocytes, NK cells, mast cells, basophils, smooth muscle cells, endothelial cells, dendritic cells, activated and resting B cells, plus activated platelets and CD4+ T cells (13, 14). CD40L ligation of CD40 on dendritic cells (DC) initiates DC maturation and differentiation. CD40L signaling into naïve B cells promotes germinal center formation and isotope switching. With IL-21, CD40L generates IgA plus IgG3; with IL-4, CD40L generates IgG1 secretion (14, 15).

References

  1. Zhang, G. (2004) Curr. Opin. Struct. Biol. 14:154.
  2. Hehlgans, T. and K. Pfeffer (2005) Immunology 115:1.
  3. Quezada, S.A. et al. (2004) Annu. Rev. Immunol. 22:307.
  4. Graf, D. et al. (1992) Eur. J. Immunol. 22:3191.
  5. Hollenbaugh, D. et al. (1992) EMBO J. 11:4313.
  6. Khandekar, S.S. et al. (2001) Prot. Exp. Purif. 23:301.
  7. Pietravalle, F. et al. (1996) J. Biol. Chem. 271:5965.
  8. Garber, E. et al. (1999) J. Biol. Chem. 274:33545.
  9. Vakkalanka, R.K. et al. (1999) Arthritis Rheum. 42:871.
  10. Eissner, G. et al. (2004) Cytokine Growth Factor Rev. 15:353.
  11. Prasad, K.S. et al. (2003) Proc. Natl. Acad. Sci. USA 100:12367.
  12. Mikolajczak, S.A. et al. (2004) J. Exp. Med. 199:1025.
  13. Lievens, D. et al. (2009) Thromb. Haemost. 102:206.
  14. Elgueta, R. et al. (2009) Immunol. Rev. 229:152.
  15. Avery, D.T. et al. (2008) J. Immunol. 181:1767.

Alternate Names

CD154, CD40L, CD40LG, gp39, TNFSF5, TRAP

Entrez Gene IDs

959 (Human); 21947 (Mouse); 84349 (Rat)

Gene Symbol

CD40LG

UniProt

Additional CD40 Ligand/TNFSF5 Products

Product Documents for Human CD40 Ligand/TNFSF5 Alexa Fluor® 405-conjugated Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human CD40 Ligand/TNFSF5 Alexa Fluor® 405-conjugated Antibody


This product is provided under an agreement between Life Technologies Corporation and R&D Systems, Inc, and the manufacture, use, sale or import of this product is subject to one or more US patents and corresponding non-US equivalents, owned by Life Technologies Corporation and its affiliates. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components (1) in manufacturing; (2) to provide a service, information, or data to an unaffiliated third party for payment; (3) for therapeutic, diagnostic or prophylactic purposes; (4) to resell, sell, or otherwise transfer this product or its components to any third party, or for any other commercial purpose. Life Technologies Corporation will not assert a claim against the buyer of the infringement of the above patents based on the manufacture, use or sale of a commercial product developed in research by the buyer in which this product or its components was employed, provided that neither this product nor any of its components was used in the manufacture of such product. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, Cell Analysis Business Unit, Business Development, 29851 Willow Creek Road, Eugene, OR 97402, Tel: (541) 465-8300. Fax: (541) 335-0354.

For research use only

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