Recombinant Human CD40 Ligand/TNFSF5 (HEK293-expressed), CF

Analyzed by SEC-MALS. HA tag
Catalog # Availability Size / Price Qty
6420-CLB-050
Recombinant Human CD40 Ligand/TNFSF5 Protein SEC-MALS.
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Citations (17)
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Recombinant Human CD40 Ligand/TNFSF5 (HEK293-expressed), CF Summary

Product Specifications

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Level
<0.10 EU per 1 μg of the protein by the LAL method.
Activity
Measured in a cell proliferation assay using human B cells. The ED50 for this effect is 0.250-3.00 ng/mL in the presence of Recombinant Human IL-4 (Catalog # 204-IL) and a cross linking HA Tag Antibody (Catalog # MAB060).
Source
Human embryonic kidney cell, HEK293-derived human CD40 Ligand/TNFSF5 protein
YPYDVPDYAGCN4-IZGGGSGGGSGGGSHuman CD40 Ligand
(Met113-Leu261)
Accession # P29965.1
N-terminusC-terminus
Accession #
N-terminal Sequence
Analysis
Tyr (of HA tag)
Predicted Molecular Mass
22 kDa
SDS-PAGE
20-27 kDa, under reducing conditions.

Product Datasheets

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6420-CLB

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

6420-CLB

Formulation Lyophilized from a 0.2 μm filtered solution in PBS and EDTA with Trehalose.
Reconstitution Reconstitute at 500 μg/mL in PBS.
Shipping The product is shipped at ambient temperature. Upon receipt, store it 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.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Scientific Data

SEC-MALS View Larger

Recombinant Human CD40 Ligand/TNFSF5 (Catalog # 6420-CLB) has a molecular weight (MW) of 66.8 kDa as analyzed by SEC-MALS, suggesting that this protein is a homotrimer.  MW may differ from predicted MW due to post-translational modifications (PTMs) present (i.e. Glycosylation).

Bioactivity View Larger

Recombinant Human CD40 Ligand/TNFSF5 (HEK293-expressed) (Catalog # 6420-CLB) stimulates the proliferation of human B cells in the presence of Recombinant Human IL‑4 (204-IL) and a cross linking HA Tag Antibody (MAB060). The ED50 for this effect is 0.250-3.00 ng/mL.

SDS-PAGE View Larger

1 μg/lane of Recombinant Human CD40 Ligand/TNFSF5 (HEK293-expressed) Protein (Catalog # 6420-CLB) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by silver staining, showing bands at 20-27 kDa.

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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Background: CD40 Ligand/TNFSF5

CD40 Ligand, also known as TNFSF, CD154, TRAP, and gp39, is a 34-39 kDa type II transmembrane glycoprotein that belongs to the TNF superfamily (1-3). Mature human CD40 Ligand consists of a 22 amino acid (aa) cytoplasmic domain, a transmembrane segment, and an 215 aa extracellular region (4, 5). The extracellular domain of human CD40 Ligand shares 74% and 76% aa sequence identity with mouse and rat CD40 Ligand, respectively. Similar to other TNF superfamily members, CD40 Ligand forms a bioactive homotrimer, both as membrane bound and soluble forms (6-9). The 18 kDa soluble form (aa 113-261) arises from proteolytic processing. Mutation and alternative splicing generate additional forms of CD40 Ligand that are often truncated or non-trimerizable (8). CD40 Ligand is expressed on platelets, as well as on activated T cells and B cells, basophils, eosinophils, fibroblasts, mast cells, monocytes, natural killer cells, vascular endothelial cells, and smooth muscle cells. CD40 Ligand binds to CD40, which is expressed on the surface of B cells, dendritic cells, macrophages, monocytes, platelets, endothelial, and epithelial cells (10). The interaction of CD40 Ligand with CD40 initiates signaling in both CD40 and CD40 Ligand expressing cells (11).  CD40 ligation by CD40 Ligand promotes B cell activation and T cell-dependent humoral responses (12, 13). CD40 Ligand dysregulation on T cells and antigen presenting cells contributes to the immune deficiency associated with HIV infection and AIDS (14, 15). It is also implicated in the pathology of multiple cardiovascular diseases including atherosclerosis, atherothrombosis, and restenosis (16, 17).

 

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) Protein Expr. 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. van Kooten, C. and J. Banchereau (1997) Curr. Opin. Immunol. 9:330.
  11. Eissner, G. et al. (2004) Cytokine Growth Factor. Rev. 15:353.
  12. Rickert, R.C. et al. (2011) Immunol. Rev. 244:115.
  13. Elgueta, R. et al. (2009) Immunol. Rev. 229:152.
  14. Kornbluth, R.S. (2000) J. Leukoc. Biol. 68:373.
  15. Chougnet, C. (2003) J. Leukoc. Biol. 74:702.
  16. Pamukcu, B. et al. (2011) Ann. Med. 43:331.
  17. Hassan, G.S. et al. (2012) Immunobiology 217:521.
Entrez Gene IDs
959 (Human); 21947 (Mouse); 84349 (Rat)
Alternate Names
CD154 antigen; CD154; CD40 antigen ligand; CD40 Ligand; CD40L; CD40-L; CD40LG; CD40LIGM; gp39; hCD40L; HIGM1; T-B cell-activating molecule; T-BAM; T-cell antigen Gp39; TNF-related activation protein; TNFSF5; TNFSF5IMD3; TRAP; TRAPtumor necrosis factor (ligand) superfamily, member 5 (hyper-IgM syndrome); tumor necrosis factor (ligand) superfamily member 5; Tumor necrosis factor ligand superfamily member 5

Citations for Recombinant Human CD40 Ligand/TNFSF5 (HEK293-expressed), CF

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.

17 Citations: Showing 1 - 10
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  1. Chromatin Landscape Underpinning Human Dendritic Cell Heterogeneity
    Authors: R Leylek, M Alcántara-, JM Granja, M Chavez, K Perez, OR Diaz, R Li, AT Satpathy, HY Chang, J Idoyaga
    Cell Rep, 2020-09-22;32(12):108180.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  2. Cholesterol metabolism drives regulatory B cell IL-10 through provision of geranylgeranyl pyrophosphate
    Authors: JA Bibby, HA Purvis, T Hayday, A Chandra, K Okkenhaug, S Rosenzweig, I Aksentijev, M Wood, HJ Lachmann, C Kemper, AP Cope, E Perucha
    Nat Commun, 2020-07-08;11(1):3412.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  3. Aspirin Reduces the Potentiating Effect of CD40L on Platelet Aggregation via Inhibition of Myosin Light Chain
    Authors: K Kojok, M Mohsen, AEH El Kadiry, W Mourad, Y Merhi
    J Am Heart Assoc, 2020-02-03;9(3):e013396.
    Species: Human
    Sample Types: Whole Cells
    Applications: Cell Culture
  4. CD24hiCD38hi and CD24hiCD27+ Human Regulatory B Cells Display Common and Distinct Functional Characteristics
    Authors: MM Hasan, L Thompson-S, G Klintmalm, AJ Demetris, J O'Leary, S Oh, H Joo
    J. Immunol., 2019-09-11;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  5. An essential role for the Zn2+ transporter ZIP7 in B cell development
    Authors: C Anzilotti, DJ Swan, B Boisson, M Deobagkar-, C Oliveira, P Chabosseau, KR Engelhardt, X Xu, R Chen, L Alvarez, R Berlinguer, KR Bull, E Cawthorne, AP Cribbs, TL Crockford, TS Dang, A Fearn, EJ Fenech, SJ de Jong, BC Lagerholm, CS Ma, D Sims, B van den Be, Y Xu, AJ Cant, G Kleiner, TR Leahy, MT de la More, JM Puck, RS Shapiro, M van der Bu, JR Chapman, JC Christians, B Davies, JA McGrath, S Przyborski, M Santibanez, SG Tangye, A Werner, GA Rutter, S Padilla-Pa, JL Casanova, RJ Cornall, ME Conley, S Hambleton
    Nat. Immunol., 2019-02-04;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  6. Condition-dependent generation of aquaporin-4 antibodies from circulating B cells in neuromyelitis optica
    Authors: R Wilson, M Makuch, AK Kienzler, J Varley, J Taylor, M Woodhall, J Palace, MI Leite, P Waters, SR Irani
    Brain, 2018-04-01;141(4):1063-1074.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  7. An anti-TL1A antibody for the treatment of asthma and inflammatory bowel disease
    Authors: AW Clarke, L Poulton, D Shim, D Mabon, D Butt, M Pollard, V Pande, J Husten, J Lyons, C Tian, AG Doyle
    MAbs, 2018-03-05;0(0):1-43.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Surface Plasmon Resonance (SPR
  8. Distinct CD40L receptors mediate inflammasome activation and secretion of IL-1? and MCP-1 in cultured human retinal pigment epithelial cells
    Authors: ZM Bian, MG Field, SG Elner, JM Kahlenberg, VM Elner
    Exp. Eye Res., 2018-02-16;170(0):29-39.
    Species: Human
    Sample Types: Whole Cells
    Applications: Cell Culture
  9. Intracellular BH3 Profiling Reveals Shifts in Antiapoptotic Dependency in Human B Cell Maturation and Mitogen-Stimulated Proliferation
    Authors: J Dai, MA Luftig
    J. Immunol., 2018-01-22;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  10. Epstein-Barr Virus Induces Adhesion Receptor CD226 (DNAM-1) Expression during Primary B-Cell Transformation into Lymphoblastoid Cell Lines
    Authors: L Grossman, C Chang, J Dai, PA Nikitin, DD Jima, SS Dave, MA Luftig
    mSphere, 2017-11-29;2(6):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  11. EZH2 inhibition by tazemetostat results in altered dependency on B-cell activation signaling�in DLBCL
    Authors: D Brach, D Johnston-B, A Drew, T Lingaraj, V Motwani, NM Warholic, I Feldman, C Plescia, JJ Smith, RA Copeland, H Keilhack, E Chan-Peneb, SK Knutson, SA Ribich, A Raimondi, MJ Thomenius
    Mol. Cancer Ther., 2017-08-23;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  12. CD40L-Dependent Pathway Is Active at Various Stages of Rheumatoid Arthritis Disease Progression
    Authors: Y Guo, AM Walsh, U Fearon, MD Smith, MD Wechalekar, X Yin, S Cole, C Orr, T McGarry, M Canavan, S Kelly, TA Lin, X Liu, SM Proudman, DJ Veale, C Pitzalis, S Nagpal
    J. Immunol., 2017-04-28;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  13. Epstein-Barr virus ensures B cell survival by uniquely modulating apoptosis at early and late times after infection
    Authors: AM Price, J Dai, Q Bazot, L Patel, PA Nikitin, R Djavadian, PS Winter, CA Salinas, AP Barry, KC Wood, EC Johannsen, A Letai, MJ Allday, MA Luftig
    Elife, 2017-04-20;6(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  14. TLR-mediated STAT3 and ERK activation controls IL-10 secretion by human B cells.
    Authors: Liu B, Cao Y, Huizinga T, Hafler D, Toes R
    Eur J Immunol, 2014-04-16;44(7):2121-9.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  15. Mitogen-induced B-cell proliferation activates Chk2-dependent G1/S cell cycle arrest.
    Authors: Nikitin, Pavel A, Price, Alexande, McFadden, Karyn, Yan, Christop, Luftig, Micah A
    PLoS ONE, 2014-01-30;9(1):e87299.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  16. Widespread potential for growth-factor-driven resistance to anticancer kinase inhibitors.
    Authors: Wilson TR, Fridlyand J, Yan Y, Penuel E, Burton L, Chan E, Peng J, Lin E, Wang Y, Sosman J, Ribas A, Li J, Moffat J, Sutherlin DP, Koeppen H, Merchant M, Neve R, Settleman J
    Nature, 2012-07-26;487(7408):505-9.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  17. B cells as a therapeutic target for IFN-beta in relapsing-remitting multiple sclerosis.
    Authors: Ramgolam VS, Sha Y, Marcus KL, Choudhary N, Troiani L, Chopra M, Markovic-Plese S
    J. Immunol., 2011-03-02;186(0):4518.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay

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