Human/Mouse CTLA-4 Antibody

Catalog # Availability Size / Price Qty
MAB386-100
MAB386-SP
Human/Mouse CTLA-4 Antibody in Western Blot
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Human/Mouse CTLA-4 Antibody Summary

Species Reactivity
Human, Mouse
Specificity
Detects human CTLA-4 in direct ELISAs. Detects human and moue CTLA-4 in Western blots.
Source
Monoclonal Mouse IgG1 Clone # 1003705
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
Chinese Hamster Ovary cell line, CHO-derived human CTLA-4
Ala37-Phe162
Accession # P16410
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
2 µg/mL
See below
Agonist Activity
Measured by its ability to to inhibit anti-CD3 antibody induced IFN-gamma secretion by human peripheral blood mononuclear cells (PBMC). The ED50 for this effect is 0.05-0.4 μg/mL.
 

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.

Data Examples

Western Blot Detection  of Human and Mouse CTLA‑4 by Western Blot. View Larger

Detection of Human and Mouse CTLA‑4 by Western Blot. Western blot shows lysates of NS0 mouse myeloma cell line transfected with with Human CTLA‑4 and mouse lymph node tissue. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human/Mouse CTLA‑4 Monoclonal Antibody (Catalog # MAB386) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for CTLA‑4 at approximately 52 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Agonist Activity Inhibition of Anti-CD3 Induced IFN-gamma  Secretion by Human/Mouse CTLA‑4 Antibody. View Larger

Inhibition of Anti-CD3 Induced IFN-gamma Secretion by Human/Mouse CTLA‑4 Antibody. Anti-Human/Mouse CTLA-4 Monoclonal Antibody (Catalog # MAB386) inhibits IFN-gamma secretion in human peripheral blood mononuclear cells (PBMC) induced by immobilized Mouse Anti-Human CD3 epsilon Monoclonal Antibody (Catalog # MAB100)coatedat 1 μg/mL (100 μL/well) in a dose-dependent manner (green line) as measured by the Human IFN-gamma Quantikine ELISA Kit (Catalog # DIF50). The ED50 is typically 0.05-0.4 μg/mL.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & 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.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

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

Reconstitution Buffers

Secondary Antibodies

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