Human CTLA-4 Alexa Fluor® 647-conjugated Antibody

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IC3251R-100UG
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Human CTLA-4 Alexa Fluor® 647-conjugated Antibody Summary

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

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.

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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 degreesC 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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