Detects human CTLA-4 in direct ELISAs and Western blots. In Western blots, approximately 25% cross-reactivity with recombinant mouse CTLA-4 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human CTLA-4 Antibody

Detection of Human CTLA‑4 by Western Blot.

Western blot shows lysates of NS0 mouse myeloma cell line either mock transfected or transfected with human CTLA-4. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human CTLA-4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-386-PB) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for CTLA-4 at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

CTLA‑4 in Human Peripheral Blood Mononuclear Cells.

CTLA-4 was detected in immersion fixed human peripheral blood mononuclear cells treated with treated with PMA and calcium ionomycin using Goat Anti-Human CTLA-4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-386-PB) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Detection of CTLA‑4 in NS0 Mouse Cell Line Co-transfected with CTLA-4 and eGFP by Flow Cytometry.

NS0 mouse myeloma cell line co-transfected with human CTLA-4 and eGFP was stained with either (A) Goat Anti-Human CTLA-4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-386-PB) or (B) Normal Goat IgG Control (Catalog # AB-108-C) followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108).

View 1 more images

Applications for Human CTLA-4 Antibody

Application

Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/10⁶ cells
Sample: NS0 mouse myeloma cell line co-transfected with human CTLA-4 and eGFP

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed human peripheral blood mononuclear cells treated with PMA and calcium ionomycin

Western Blot

0.5 µg/mL
Sample: NS0 mouse myeloma cell line transfected with human CTLA-4

Reviewed Applications

Read 2 reviews rated 5 using AF-386-PB in the following applications:

Immunocytochemistry (1 Review)
Immunofluorescence (1 Review)

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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.

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 and CD28, together with their ligands B7-1 and B7-2, constitute one of the dominant costimulatory pathways that regulate T- and B-cell responses. CTLA-4 and CD28 are structurally homologous molecules that are members of the immunoglobulin (Ig) gene superfamily. Both CTLA-4 and CD28 are composed of a single
Ig V‑like extracellular domain, a transmembrane domain and an intracellular domain. CTLA-4 and CD28 are both expressed on the cell surface as disulfide-linked homodimers or as monomers. The genes encoding these two molecules are closely linked on human chromosome 2. CTLA-4 was originally identified as a gene that was specifically expressed by cytotoxic T lymphocytes. However, CTLA-4 transcripts have since been found in both Th1 and Th2, and CD4⁺and CD8⁺T cell clones. Whereas CD28 expression is constitutive on the surfaces of 95% of CD4⁺ T cells and 50% of CD8⁺T cells and is down regulated upon T cell activation, CTLA-4 expression is upregulated rapidly following T cell activation and peaks approximately 24 hours following activation. Although both CTLA-4 and CD28 can bind to the same ligands, CTLA-4 binds to B7-1 and B7-2 with 20‑100‑fold higher affinity than CD28. The physiological role of CTLA-4 in T cell costimulation is currently being studied.

References

Lenschow, D.J. et al. (1996) Annu. Rev. Immunol. 14:233.
Hathcock, K.S. and R.J. Hodes (1996) Advances in Immunol. 62:131.
Ward, S.G. (1996) Biochem. J. 318:361.

Long Name

Cytotoxic T-lymphocyte-associated Molecule 4

Alternate Names

CD152, CTLA4

Entrez Gene IDs

1493 (Human); 12477 (Mouse); 403696 (Canine); 102115124 (Cynomolgus Monkey)

Gene Symbol

CTLA4

UniProt

Q6GR94

Additional CTLA-4 Products

Product Documents for Human CTLA-4 Antibody

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Human CTLA-4 Antibody

For research use only

Citations
Reviews

Calculators
Flow Cytometry

Human CTLA-4 Antibody

R&D Systems, part of Bio-Techne | Catalog # AF-386-PB

Key Product Details

Validated by

Species Reactivity

Validated:

Cited:

Applications