Human CTLA-4 Alexa Fluor® 647-conjugated Antibody

    
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  • Species Reactivity
    Human
  • Specificity
    Detects human CTLA-4 in direct ELISAs.
  • Source
    Monoclonal Mouse IgG2B Clone # 922101
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    Chinese hamster ovary cell line CHO-derived recombinant human CTLA-4
    Ala37-Phe162
    Accession # P16410
  • Formulation
    Supplied 0.2 mg/mL in a saline solution containing BSA and Sodium Azide.
  • Label
    Alexa Fluor 647
Product Datasheets

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Applications
  •  
    Recommended
    Concentration
    Sample
  • Intracellular Staining by Flow Cytometry
    0.25-1 µg/106 cells
    Human peripheral blood mononuclear cells (PBMCs) treated with PHA were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005).
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Preparation and Storage
  • Shipping
    The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
  • Stability & Storage
    Protect from light. Do not freeze.
    • 12 months from date of receipt, 2 to 8 °C as supplied.
Background: CTLA-4

CTLA-4 (cytotoxic T-lymphocyte associated protein‑4, designated CD152), is a type I transmembrane T cell inhibitory molecule that is a member of the Ig superfamily (1, 2). Human or mouse CTLA-4 cDNA encodes 223 amino acids (aa) including a 35 aa signal sequence, a 126 aa extracellular domain (ECD) with one Ig-like V-type domain, a 21 aa transmembrane (TM) sequence, and a 41 aa cytoplasmic sequence. It is found as a covalent homodimer of 41-43 kDa (2) Within the ECD, human CTLA-4 shares 68%, 71% and 83‑86% aa sequence identity with mouse, rat and porcine/bovine/rabbit/feline/canine CTLA-4, respectively. A 174 aa form that lacks TM and cytoplasmic sequences (sCTLA-4) is possibly secreted (3-5). Isoforms of 56-79 aa that mainly contain parts of the cytoplasmic domain are reported. In mouse, an isoform lacking the Ig-like domain has ligand-independent inhibitory activity and is termed liCTLA-4 (6). CD28, which is structurally related to CTLA-4, is constitutively expressed on naïve T cells and promotes T cell activation when engaged by B7-2 on antigen-presenting cells (APC) within the immunological synapse (IS) (1, 7, 8). In contrast, CTLA-4 is recruited from intracellular vesicles to the IS beginning 1-2 days after T cell activation (2, 7, 8). It forms a linear lattice with B7-1 on APC, inducing negative regulatory signals and ending T cell activation (9). Abatacept, a therapeutic human CTLA-4-Ig fusion protein (trade name Orencia), competes with CD28 for B7-1 and B7-2 binding and has been used to antagonize T cell activation in autoimmune conditions and to enhance transplant survival (10). Mice deleted for CTLA-4 show no abnormalities until after birth, but then develop lethal autoimmune reactions due to continued T cell activation and poor control by regulatory T cells, which constitutively express CTLA-4 in wild-type mice and humans (11-13).

  • References:
    1. Harper, K. et al. (1991) J. Immunol. 147:1037.
    2. Teft, W.A. et al. (2006) Annu. Rev. Immunol. 24:65.
    3. Magistrelli, G. et al. (1999) Eur. J. Immunol. 29:3596.
    4. Tector, M. et al. (2009) BMC Immunol. 10:51.
    5. Oaks, M.K. and K.M. Hallett (2000) J. Immunol. 164:5015.
    6. Vijayakrishnan, L. et al. (2004) Immunity 20:563.
    7. Pentcheva-Hoang, T. et al. (2004) Immunity 21:401.
    8. Jansson, A. et al. (2005) J. Immunol 175:1575.
    9. Darlington, P.J. et al. (2005) J. Immunol. 175:996.
    10. Platt, A.M. et al. (2010) J. Immunol. 185:1558.
    11. Wing, K. et al. (2008) Science 322:271.
    12. Friedline, R.H. et al. (2009) J. Exp. Med. 206:421.
    13. Jain, N. et al. (2010) Proc. Natl. Acad. Sci. USA 107:1524.
  • Long Name:
    Cytotoxic T-lymphocyte-associated Molecule 4
  • Entrez Gene IDs:
    1493 (Human); 12477 (Mouse); 403696 (Canine); 102115124 (Cynomolgus Monkey)
  • Alternate Names:
    CD; CD152 antigen; CD152; CD152IDDM12; CD28; celiac disease 3; CELIAC3; CTLA4; CTLA-4; cytotoxic T-lymphocyte antigen 4; cytotoxic T-lymphocyte protein 4; Cytotoxic T-lymphocyte-associated antigen 4; cytotoxic T-lymphocyte-associated protein 4; cytotoxic T-lymphocyte-associated serine esterase-4; GRD4; GSE; ICOS; ligand and transmembrane spliced cytotoxic T lymphocyte associated antigen 4
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Isotype Controls
Description Application Cat# Citations Images  

Mouse IgG2B Alexa Fluor® 647-conjugated Isotype Control

Ctrl IC0041R
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Staining Reagents
Description Application Cat# Citations Images  

Flow Cytometry Permeabilization/Wash Buffer I (1X)

Flow FC005 5
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Flow Cytometry Fixation Buffer (1X)

Flow FC004 3
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Flow Cytometry Fixation & Permeabilization Buffer Kit I

Flow FC009 4
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Flow Cytometry Fixation/Permeabilization Buffer I (1X)

Flow FC007 2
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