Recombinant Mouse CD40 Ligand/TNFSF5 (HA-tag) Protein

Recombinant Mouse CD40 Ligand/TNFSF5 (Catalog # 8230-CL) has a molecular weight (MW) of 66.9 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).

• Reactivity: Mu
• Applications: Bioactivity

Recombinant Mouse CD40 Ligand/TNFSF5 (HA-tag) Protein Summary

• Additional Information: Analyzed by SEC-MALS
• Details of Functionality: Measured in a cell proliferation assay using mouse splenic B cells in the presence of IL-4. Banchereau, J. et al. (1991) Science 251:70. The ED₅₀ for this effect is 0.0700-0.350 ng/mL in the presence of a cross‑linking antibody, Mouse Anti-Hemagglutinin/HA Peptide Monoclonal Antibody (Catalog # MAB060).
• Source: Chinese Hamster Ovary cell line, CHO-derived mouse CD40 Ligand/TNFSF5 protein
  

  HA (YPYDVPDYA)	GCN4-IZ	GGGSGGGSGGGS	Mouse CD40 Ligand/TNFSF5 (Met112-Leu260) Accession # P27548
  N-terminus			C-terminus

• Accession #: P27548
• N-terminal Sequence: Tyr
• Protein/Peptide Type: Recombinant Proteins
• Gene: Cd40lg
• Purity: >95%, by SDS-PAGE with silver staining.
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 22 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 24-29 kDa, reducing conditions

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.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS and NaCl with BSA as a carrier protein.
• Purity: >95%, by SDS-PAGE with silver staining.
• Reconstitution Instructions: Reconstitute at 100 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin.

Notes

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

Alternate Names for Recombinant Mouse CD40 Ligand/TNFSF5 (HA-tag) Protein

• 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

Background

CD40 Ligand, also known as TNFSF5, CD154, TRAP, or gp39, is a 33-39 kDa type II transmembrane glycoprotein member of the TNF superfamily (1, 2). Mature mouse CD40 Ligand consists of a 22 amino acid (aa) cytoplasmic domain, a transmembrane segment, and a 214 aa extracellular region (1). The extracellular domain of mouse CD40 Ligand shares 75% and 93% amino acid (aa) sequence identity with the human and rat proteins, respectively. CD40 Ligand is expressed as a homotrimer on platelets and activated T cells and B cells. It is up-regulated following stimulation of basophils, eosinophils, fibroblasts, mast cells, monocytes, natural killer cells, vascular endothelial cells, and smooth muscle cells. CD40 Ligand binds and activates CD40, which is expressed on the surface of B cells, dendritic cells, macrophages, monocytes, platelets, endothelial cells, and epithelial cells (3). The 18 kDa soluble form (aa 112-260)  arises from proteolytic processing and retains the ability to bind and activate CD40 (4, 5). Monomeric, dimeric, and trimeric forms of soluble CD40 Ligand bind to oligomeric CD40 on cell membranes (2). CD40 ligation by CD40 Ligand promotes B cell activation and T cell-dependent humoral responses (6, 7). CD40 Ligand dysregulation on T cells and antigen presenting cells contributes to the immune deficiency associated with HIV infection and AIDS (8, 9). It is also implicated in the pathology of multiple cardiovascular diseases including atherosclerosis, atherothrombosis, and restenosis (10, 11).

1. Armitage, R.J. et al. (1992) Nature 357:80.
2. Hollenbaugh, D. et al. (1992) EMBO J. 11:4313.
3. van Kooten, C. and J. Banchereau (1997) Curr. Opin. Immunol. 9:330.
4. Graf, D. et al. (1995) Eur. J. Immunol. 25:1749.
5. Mazzei, G.J. et al. (1995) J. Biol. Chem. 270:7025.
6. Rickert, R.C. et al. (2011) Immunol. Rev. 244:115.
7. Elgueta, R. et al. (2009) Immunol. Rev. 229:152.
8. Kornbluth, R.S. (2000) J. Leukoc. Biol. 68:373.
9. Chougnet, C. (2003) J. Leukoc. Biol. 74:702.
10. Pamukcu, B. et al. (2011) Ann. Med. 43:331.
11. Hassan, G.S. et al. (2012) Immunobiology 217:521.

Publications for CD40 Ligand/TNFSF5 (8230-CL)(20)

Publications using 8230-CL

• A Di Pietro, J Polmear, L Cooper, T Damelang, T Hussain, L Hailes, K O'Donnell, V Udupa, T Mi, S Preston, A Shtewe, U Hershberg, SJ Turner, NL La Gruta, AW Chung, DM Tarlinton, CD Scharer, KL Good-Jacob Targeting BMI-1 in B cells restores effective humoral immune responses and controls chronic viral infection Nature Immunology, 2021-11-29;0(0):. 2021-11-29 [PMID: 34845392] (Bioassay, Mouse)
• L Kealy, A Di Pietro, L Hailes, S Scheer, L Dalit, JR Groom, C Zaph, KL Good-Jacob The Histone Methyltransferase DOT1L Is Essential for Humoral Immune Responses Cell Reports, 2020-12-15;33(11):108504. 2020-12-15 [PMID: 33326791] (Bioassay, Transgenic Mouse)
• K Abdi, K Laky, K Padhan, C Petrovas, J Skinner, J Kabat, DW Dorward, J Brzostowsk, EO Long, G Trinchieri, R Varma Cutting Edge: Quantitative Determination of CD40L Threshold for IL-12 and IL-23 Production from Dendritic Cells J. Immunol., 2018-10-12;0(0):. 2018-10-12 [PMID: 30315139] (Cell Culture)
• MS Russell, A Muralidhar, L Larocque, J Cao, Y Deschambau, J Varga, SN Thulasi Ra, X Li Identification and characterisation of the CD40-ligand of Sigmodon hispidus PLoS ONE, 2018-07-27;13(7):e0199067. 2018-07-27 [PMID: 30052641] (ELISA Developmet, Mouse)
• S Trezise, A Karnowski, PL Fedele, S Mithraprab, Y Liao, K D'Costa, AJ Kueh, MP Hardy, CM Owczarek, MJ Herold, A Spencer, W Shi, SN Willis, SL Nutt, LM Corcoran Mining the Plasma Cell Transcriptome for Novel Cell Surface Proteins Int J Mol Sci, 2018-07-24;19(8):. 2018-07-24 [PMID: 30042348] (Bioassay, Mouse)
• BG Barwick, CD Scharer, RJ Martinez, MJ Price, AN Wein, RR Haines, APR Bally, JE Kohlmeier, JM Boss B cell activation and plasma cell differentiation are inhibited by de novo DNA methylation Nat Commun, 2018-05-15;9(1):1900. 2018-05-15 [PMID: 29765016] (Bioassay, Mouse)
• A Kumar, A Priya, T Ahmed, C Grundström, N Negi, T Grundström Regulation of the DNA Repair Complex during Somatic Hypermutation and Class-Switch Recombination J. Immunol., 2018-05-04;0(0):. 2018-05-04 [PMID: 29728513] (Bioassay, Mouse)
• CD Scharer, BG Barwick, M Guo, APR Bally, JM Boss Plasma cell differentiation is controlled by multiple cell division-coupled epigenetic programs Nat Commun, 2018-04-27;9(1):1698. 2018-04-27 [PMID: 29703886] (Bioassay, Mouse)
• K Ochiai, H Kondo, Y Okamura, H Shima, Y Kurokochi, K Kimura, R Funayama, T Nagashima, K Nakayama, K Yui, K Kinoshita, K Igarashi Zinc finger-IRF composite elements bound by Ikaros/IRF4 complexes function as gene repression in plasma cell Blood Adv, 2018-04-24;2(8):883-894. 2018-04-24 [PMID: 29669755] (Bioassay, Mouse)
• W Ise, K Fujii, K Shiroguchi, A Ito, K Kometani, K Takeda, E Kawakami, K Yamashita, K Suzuki, T Okada, T Kurosaki T Follicular Helper Cell-Germinal Center B Cell Interaction Strength Regulates Entry into Plasma Cell or Recycling Germinal Center Cell Fate Immunity, 2018-04-17;48(4):702-715.e4. 2018-04-17 [PMID: 29669250] (Bioassay, Mouse)
• Y Miura, M Morooka, N Sax, R Roychoudhu, A Itoh-Nakad, A Brydun, R Funayama, K Nakayama, S Satomi, M Matsumoto, K Igarashi, A Muto Bach2 Promotes B Cell Receptor-Induced Proliferation of B Lymphocytes and Represses Cyclin-Dependent Kinase Inhibitors J. Immunol., 2018-03-14;0(0):. 2018-03-14 [PMID: 29540581] (Bioassay, Mouse)
• S Cosenza, K Toupet, M Maumus, P Luz-Crawfo, O Blanc-Brud, C Jorgensen, D Noël Mesenchymal stem cells-derived exosomes are more immunosuppressive than microparticles in inflammatory arthritis Theranostics, 2018-02-03;8(5):1399-1410. 2018-02-03 [PMID: 29507629] (Bioassay, Mouse)
• M Burbage, SJ Keppler, B Montaner, PK Mattila, FD Batista The Small Rho GTPase TC10 Modulates B Cell Immune Responses J. Immunol., 2017-07-26;0(0):. 2017-07-26 [PMID: 28747344] (Bioassay, Mouse)
• BC Lee, SG Lee, MK Choo, JH Kim, HM Lee, S Kim, DE Fomenko, HY Kim, JM Park, VN Gladyshev Selenoprotein MsrB1 promotes anti-inflammatory cytokine gene expression in macrophages and controls immune response in vivo Sci Rep, 2017-07-11;7(1):5119. 2017-07-11 [PMID: 28698597] (Bioassay, Mouse)
• TLR3-Induced Maturation of Murine Dendritic Cells Regulates CTL Responses by Modulating PD-L1 Trafficking PLoS ONE, 2016-12-02;11(12):e0167057. 2016-12-02 [PMID: 27911948] (Bioassay, Mouse)
• Thomas Grundström Regulated localization of an AID complex with E2A, PAX5 and IRF4 at the Igh locus Mol. Immunol., 2016-11-12;80(0):78-90. 2016-11-12 [PMID: 27835756] (Bioassay, Mouse)
• Ackermann J, Nys J, Schweighoffer E, McCleary S, Smithers N, Tybulewicz V Syk tyrosine kinase is critical for B cell antibody responses and memory B cell survival. J Immunol, 2015-04-10;194(10):4650-6. 2015-04-10 [PMID: 25862820] (Bioassay, Mouse)
• Jacque E, Schweighoffer E, Visekruna A, Papoutsopoulou S, Janzen J, Zillwood R, Tarlinton D, Tybulewicz V, Ley S IKK-induced NF-kappaB1 p105 proteolysis is critical for B cell antibody responses to T cell-dependent antigen. J Exp Med, 2014-09-15;211(10):2085-101. 2014-09-15 [PMID: 25225457] (Bioassay, Mouse)
• Song Z, Zhu X, Jin R, Wang C, Yan J, Zheng Q, Nanda A, Granger D, Li G Roles of the kinase TAK1 in CD40-mediated effects on vascular oxidative stress and neointima formation after vascular injury. PLoS ONE, 2014-07-22;9(7):e101671. 2014-07-22 [PMID: 25050617] (Bioassay, Mouse)
• Figgett W, Fairfax K, Vincent F, Le Page M, Katik I, Deliyanti D, Quah P, Verma P, Grumont R, Gerondakis S, Hertzog P, O'Reilly L, Strasser A, Mackay F The TACI receptor regulates T-cell-independent marginal zone B cell responses through innate activation-induced cell death. Immunity, 2013-09-05;39(3):573-83. 2013-09-05 [PMID: 24012421] (Bioassay, Mouse)

FAQs for CD40 Ligand/TNFSF5 (8230-CL). (Showing 1 - 1 of 1 FAQs).

1. I am looking for paired ABs for use in detecting CD40L (rat) by ELISA. Do you have such a reagent(s) available?
   • Unfortunately we currently do not have any CD40 Ligand antibody pairs that have been validated in Rat. The only antibody pairs we offer for CD40L are validated in Human. I apologize for this inconvenience. If you would be interested in testing one of our products in Rat I'd like to invite you to participate in our Innovators Reward Program. Please contact us at innovators@novusbio.com with any questions regarding this program.

Bioinformatics

• Gene Symbol: Cd40lg
• Uniprot:
  • P27548()