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Mouse Monoclonal to CD4

MEM-241 (IgG1)

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Background

CD4 (T4) is a single chain transmembrane glycoprotein and belongs to immunoglobulin supergene family. In extracellular region there are 4 immunoglobulin-like domains (1 Ig-like V-type and 3 Ig-like C2-type). Transmembrane region forms 25 aa, cytoplasmic tail consists of 38 aa. Domains 1,2 and 4 are stabilized by disulfide bonds. The intracellular domain of CD4 is associated with p56Lck, a Src-like protein tyrosine kinase. It was described that CD4 segregates into specific detergent-resistant T-cell membrane microdomains.
Extracellular ligands: MHC class II molecules (binds to CDR2-like region in CD4 domain 1); HIV envelope protein gp120 (binds to CDR2-like region in CD4 domain 1); IL-16 (binds to CD4 domain 3), Human seminal plasma glycoprotein gp17 (binds to CD4 domain 1), L-selectin
Intracellular ligands: p56Lck
CD4 is a co-receptor involved in immune response (co-receptor activity in binding to MHC class II molecules) and HIV infection (human immunodeficiency virus; CD4 is primary receptor for HIV-1 surface glycoprotein gp120). CD4 regulates T-cell activation, T/B-cell adhesion, T-cell diferentiation, T-cell selection and signal transduction. Defects in antigen presentation (MHC class II) cause dysfunction of CD4+ T-cells and their almost complete absence in patients blood, tissue and organs (SCID immunodeficiency).

Specificity

The antibody MEM-241 recognizes CD4 antigen, a 55 kDa transmebrane glycoprotein expressed on a subset of T lymphocytes (“helper“ T-cells) and also on monocytes, tissue macrophages and granulocytes.

HCDM (former HLDA VIII) Meeting, May 2006, Québec, Canada; WS Code M241

Regulatory Status

Immunogen

2 N-terminal domains of human CD4 fused to human IgG1 Fc

Species Reactivity:

  • Human
  • Other not tested

Negative Species:

Applications:

  • Flow Cytometry
    Recommended dilution:1 μg/ml
    Positive control:peripheral blood, HPB T cell line
    Application note:excellent antibody for FC application
  • Western Blotting
    Recommended dilution: 1-2 μg/ml
    Positive control: peripheral blood, HPB T cell line
    Sample preparation: Resuspend approx. 50 mil. cells in 1 ml cold Lysis buffer (1% laurylmaltoside in 20 mM Tris/Cl, 100 mM NaCl pH 8.2, 50 mM NaF including Protease inhibitor Cocktail). Incubate 60 min on ice. Centrifuge to remove cell debris. Mix lysate with non-reducing SDS-PAGE sample buffer.
    Application note: Non-reducing conditions.
Usage note:
Indicated dilutions are recommended starting points for use of this product. Working concentrations should be determined by the investigator.

General references

  • *Millan J, Cerny J, Horejsi V, Alonso MA: CD4 segregates into specific detergent-resistant T-cell membrane microdomains. Tissue Antigens. 1999 Jan;53(1):33-40. [Abstract]
  • *Foti M, Phelouzat MA, Holm A, Rasmusson BJ, Carpentier JL: p56Lck anchors CD4 to distinct microdomains on microvilli. Proc Natl Acad Sci U S A. 2002 Feb 19;99(4):2008-13. [Abstract] [Full Text]
  • Clapham PR, McKnight A: Cell surface receptors, virus entry and tropism of primate lentiviruses. J Gen Virol. 2002 Aug;83(Pt 8):1809-29. [Abstract] [Full Text]
  • Product Specific References

  • *Brdickova N. et al.: LIME: a new membrane Raft-associated adaptor protein involved in CD4 and CD8 coreceptor signaling. J Exp Med. 2003 Nov 17;198(10):1453-62. [Abstract] [Full Text]
  • *Zola H, Swart B, Banham A, Barry S, Beare A, Bensussan A, Boumsell L, D Buckley C, Buhring HJ, Clark G, Engel P, Fox D, Jin BQ, Macardle PJ, Malavasi F, Mason D, Stockinger H, Yang X: CD molecules 2006--human cell differentiation molecules. J Immunol Methods. 2007 Jan 30;319(1-2):1-5. [Abstract]
  • *Karlsson KR, Cowley S, Martinez FO, Shaw M, Minger SL, James W: Homogeneous monocytes and macrophages from human embryonic stem cells following coculture-free differentiation in M-CSF and IL-3. Exp Hematol. 2008 Sep;36(9):1167-75. [Abstract] [Full Text]
  • *Manasa J, Musabaike H, Masimirembwa C, Burke E, Luthy R, Mudzori J: Evaluation of the Partec flow cytometer against the BD FACSCalibur system for monitoring immune responses of human immunodeficiency virus-infected patients in Zimbabwe. Clin Vaccine Immunol. 2007 Mar;14(3):293-8. [Abstract] [Full Text]
  • *Anderson AE, Sayers BL, Haniffa MA, Swan DJ, Diboll J, Wang XN, Isaacs JD, Hilkens CM: Differential regulation of naïve and memory CD4+ T cells by alternatively activated dendritic cells. J Leukoc Biol. 2008 Jul;84(1):124-33. [Abstract] [Full Text]
  • *Hovden AO, Karlsen M, Jonsson R, Aarstad HJ, Appel S: Maturation of monocyte derived dendritic cells with OK432 boosts IL-12p70 secretion and conveys strong T-cell responses. BMC Immunol. 2011 Jan 5;12:2. [Abstract] [Full Text]
  • For research use only. Not for drug, diagnostic or other use.

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  • Example Data


    MEM-241
    Fig. 1


    Fig. 1. Surface staining of human peripheral blood cells with anti-human CD4 (MEM-241) FITC.




    MEM-241 MC
    Fig. 2

    Fig. 2. Surface staining (mass cytometry) of PBMC after Ficoll-Paque separation with anti-human CD4 (MEM-241) Er166. Gated on singlets.



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