Human/Mouse CTLA-4 Antibody

Catalog #: MAB386 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB386-100
MAB386-SP

Detection of Human and Mouse CTLA‑4 by 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 either lyophilized or 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.

Scientific Data

Western Blot

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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

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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 CD3e Monoclonal Antibody (Catalog # MAB100) coated at 1 µg/mL (100 µL/well) in a dose-dependent manner (green line) as measured by the Human IFN-gamma Quantikine ELISA Kit (Catalog # DIF50C). The ED₅₀ is typically 0.05-0.4 µg/mL.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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

Harper, K. et al. (1991) J. Immunol. 147:1037.
Teft, W.A. et al. (2006) Annu. Rev. Immunol. 24:65.
Magistrelli, G. et al. (1999) Eur. J. Immunol. 29:3596.
Tector, M. et al. (2009) BMC Immunol. 10:51.
Oaks, M.K. and K.M. Hallett (2000) J. Immunol. 164:5015.
Vijayakrishnan, L. et al. (2004) Immunity 20:563.
Pentcheva-Hoang, T. et al. (2004) Immunity 21:401.
Jansson, A. et al. (2005) J. Immunol 175:1575.
Darlington, P.J. et al. (2005) J. Immunol. 175:996.
Platt, A.M. et al. (2010) J. Immunol. 185:1558.
Wing, K. et al. (2008) Science 322:271.
Friedline, R.H. et al. (2009) J. Exp. Med. 206:421.
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