Human CTLA-4 Antibody

Catalog # Availability Size / Price Qty
MAB3253-100
MAB3253-SP
Detection of CTLA‑4 in NS0 Mouse Cell Line Transfected with Human CTLA-4 and eGFP by Flow Cytometry.
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Human CTLA-4 Antibody Summary

Species Reactivity
Human
Specificity
Detects human CTLA-4 in direct ELISAs.
Source
Recombinant Monoclonal Rabbit IgG Clone # 2188B
Purification
Protein A or G purified from cell culture supernatant
Immunogen
Chinese hamster ovary cell line CHO-derived recombinant 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
Flow Cytometry
0.25 µg/106 cells
See below
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 
Immunocytochemistry
8-25 µg/mL
See below

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

Flow Cytometry Detection of CTLA‑4 in NS0 Mouse Cell Line Transfected with Human CTLA-4 and eGFP by Flow Cytometry. View Larger

Detection of CTLA‑4 in NS0 Mouse Cell Line Transfected with Human CTLA-4 and eGFP by Flow Cytometry. NS0 mouse myeloma cell line transfected with human CTLA-4 and eGFP was stained with (A) Rabbit Anti-Human CTLA‑4 Monoclonal Antibody (Catalog # MAB3253) or (B) Normal Rabbit IgG Control Antibody (Catalog # MAB1050) followed by APC-conjugated Goat anti-Rabbit IgG Secondary Antibody (Catalog # F0111). View our protocol for Staining Membrane-associated Proteins.

Immunocytochemistry CTLA‑4  in Human PBMCs. View Larger

CTLA‑4 in Human PBMCs. CTLA‑4 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) stimulated with PHA using Rabbit Anti-Human CTLA‑4 Monoclonal Antibody (Catalog # MAB3253) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

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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Cell and Tissue Staining Kits

Isotype Controls

Normal Rabbit IgG Control

Ctrl,CyTOF-ready

Reconstitution Buffers

Secondary Antibodies

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