Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human, Rat, Xenograft

Applications

Validated:

Western Blot, Flow Cytometry, Immunocytochemistry, CyTOF-ready

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Flow Ctyometry, In vivo assay

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all CTLA-4 Primary Antibodies »

Product Specifications

Immunogen

S. frugiperda insect ovarian cell line Sf 21-derived recombinant human CTLA-4
Ala37-Phe162
Accession # Q6GR94

Specificity

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 antibody by Western Blot.

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 antibody in Human Peripheral Blood Mononuclear Cells by Immunocytochemistry (ICC).

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

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).

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:

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Flow Cytometry

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.

Calculators

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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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

  1. Lenschow, D.J. et al. (1996) Annu. Rev. Immunol. 14:233.
  2. Hathcock, K.S. and R.J. Hodes (1996) Advances in Immunol. 62:131.
  3. 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

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human CTLA‑4 Antibody

For research use only

Citations for Human CTLA‑4 Antibody

Customer Reviews for Human CTLA‑4 Antibody (2)

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2 Customer Ratings
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  • Human CTLA-4 Antibody
    Name: Anonymous
    Application: Immunocytochemistry
    Sample Tested: Breast cancer tissue and Adult lung
    Species: Human
    Verified Customer | Posted 06/21/2017
  • Name: Christopher England
    Application: Immunofluorescence
    Sample Tested: Human lung cancer cell line MiaPaCa-2
    Species: Human
    Verified Customer | Posted 08/05/2016
    CTLA-4 Expression in Pancreatic Cancer Cell Line (MiaPaCa-2)
    Human CTLA‑4 Antibody AF-386-PB

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Protocols

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