Human CTLA-4 Antibody AF-386-PB: R&D Systems

Human CTLA-4 Antibody

(2 citations)   
  • Species Reactivity
    Human
  • 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.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    S. frugiperda insect ovarian cell line Sf 21-derived recombinant human CTLA-4
    Ala37-Phe162
    Accession # Q6GR94
  • 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
    0.5 µg/mL
    See below
  • 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
    5-15 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
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.
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).
Immunocytochemistry
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.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • 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 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
  • 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
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
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Species
Applications
Sample Type
  1. PD-L1 expression and its relationship with oncogenic drivers in non-small cell lung cancer (NSCLC)
    Authors: L Jiang, X Su, T Zhang, X Yin, M Zhang, H Fu, H Han, Y Sun, L Dong, J Qian, Y Xu, X Fu, PR Gavine, Y Zhou, K Tian, J Huang, D Shen, H Jiang, Y Yao, B Han, Y Gu
    Oncotarget, 2017;8(16):26845-26857.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC
  2. Inhibitory receptors are expressed by Trypanosoma cruzi-specific effector T cells and in hearts of subjects with chronic Chagas disease.
    Authors: Arguello RJ, Albareda MC, Alvarez MG, Bertocchi G, Armenti AH, Vigliano C, Meckert PC, Tarleton RL, Laucella SA
    PLoS ONE, 2012;7(5):e35966.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
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Donkey Anti-Goat IgG NorthernLights™ NL637-conjugated Antibody

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Chicken Anti-Goat IgG Biotinylated Antibody

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Rabbit Anti-Goat IgG Biotinylated Antibody

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Rabbit Anti-Goat IgG (H+L) Affinity Purified PAb, X Absorbed

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Donkey Anti-Goat IgG (H+L) PerCP-conjugated Antibody

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