| Clone: | MEM-241 |
|---|---|
| Isotype: | Mouse IgG1 |
| 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 |
| Immunogen: | 2 N-terminal domains of human CD4 fused to human IgG1 Fc |
| Species Reactivity: | Human, Other not tested |
| Application: |
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. |
| Purity: | > 95% (by SDS-PAGE) |
| Purification: | Purified from ascites by protein-A affinity chromatography. |
| Concentration: | 1 mg/ml |
| Storage Buffer: | Phosphate buffered saline (PBS) with 15 mM sodium azide, approx. pH 7.4 |
| Storage / Stability: | Store at 2-8°C. Do not use after expiration date stamped on vial label. For long-term storage aliquot and store at -20°C. Avoid freeze/thaw cycles. |
| Expiration: | See vial label |
| Lot Number: | See vial label |
| 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). |
| 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.
*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.
Clapham PR, McKnight A: Cell surface receptors, virus entry and tropism of primate lentiviruses. J Gen Virol. 2002 Aug;83(Pt 8):1809-29.
*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.
*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.
*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.
*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.
*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.
*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.
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