Recombinant

Recombinant Anti-CTLA4 antibody [CAL49] (ab237712)

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Overview

  • Product name

    Anti-CTLA4 antibody [CAL49]
    See all CTLA4 primary antibodies

  • Description

    Rabbit monoclonal [CAL49] to CTLA4

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, WB, Flow Cyt, IPmore details
    Unsuitable for: ICC/IF

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    Synthetic peptide. This information is considered to be commercially sensitive.

  • Positive control

    • IHC-P: Human tonsil, lymph node and breast carcinoma tissues. WB: Human PBMCs (treated with 10µg/ml PHA for 2 days) whole cell lysate; mouse splenocytes (treated with 2.5µg/ml Concanavalin A (ConA) for 3 days) whole cell lysate. Flow cyt: Human PBMCs (treated with 10µg/ml PHA for 2 days); mouse splenocytes (treated with 2.5µg/ml Concanavalin A (ConA) for 3 days). IP: Human tonsil lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab237712 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 25 kDa (predicted molecular weight: 25 kDa).
Flow Cyt 1/400.
IP 1/30.
  • Application notes
    Is unsuitable for ICC/IF.

    • Target

    • Function

      Inhibitory receptor acting as a major negative regulator of T-cell responses. The affinity of CTLA4 for its natural B7 family ligands, CD80 and CD86, is considerably stronger than the affinity of their cognate stimulatory coreceptor CD28.

    • Tissue specificity

      Widely expressed with highest levels in lymphoid tissues. Detected in activated T-cells where expression levels are 30- to 50-fold less than CD28, the stimulatory coreceptor, on the cell surface following activation.

    • Involvement in disease

      Genetic variation in CTLA4 influences susceptibility to systemic lupus erythematosus (SLE) [MIM:152700]. SLE is a chronic, inflammatory and often febrile multisystemic disorder of connective tissue. It affects principally the skin, joints, kidneys and serosal membranes. SLE is thought to represent a failure of the regulatory mechanisms of the autoimmune system.
      Note=Genetic variations in CTLA4 may influence susceptibility to Graves disease, an autoimmune disorder associated with overactivity of the thyroid gland and hyperthyroidism.
      Genetic variation in CTLA4 is the cause of susceptibility to diabetes mellitus insulin-dependent type 12 (IDDM12) [MIM:601388]. A multifactorial disorder of glucose homeostasis that is characterized by susceptibility to ketoacidosis in the absence of insulin therapy. Clinical fetaures are polydipsia, polyphagia and polyuria which result from hyperglycemia-induced osmotic diuresis and secondary thirst. These derangements result in long-term complications that affect the eyes, kidneys, nerves, and blood vessels.
      Genetic variation in CTLA4 is the cause of susceptibility to celiac disease type 3 (CELIAC3) [MIM:609755]. It is a multifactorial disorder of the small intestine that is influenced by both environmental and genetic factors. It is characterized by malabsorption resulting from inflammatory injury to the mucosa of the small intestine after the ingestion of wheat gluten or related rye and barley proteins. In its classic form, celiac disease is characterized in children by malabsorption and failure to thrive.

    • Sequence similarities

      Contains 1 Ig-like V-type (immunoglobulin-like) domain.

    • Post-translational
      modifications

      N-glycosylation is important for dimerization.
      Phosphorylation at Tyr-201 prevents binding to the AP-2 adapter complex, blocks endocytosis, and leads to retention of CTLA4 on the cell surface.

    • Cellular localization

      Cell membrane. Exists primarily an intracellular antigen whose surface expression is tightly regulated by restricted trafficking to the cell surface and rapid internalisation and.
    • Information by UniProt

    • Database links

      • Alternative names

        • ALPS5 antibody
        • CD 152 antibody
        • CD antibody
        • CD152 antibody
        • CD152 antigen antibody
        • CD152 isoform antibody
        • Celiac disease 3 antibody
        • CELIAC3 antibody
        • CTLA 4 antibody
        • CTLA-4 antibody
        • CTLA4 antibody
        • CTLA4_HUMAN antibody
        • Cytotoxic T cell associated 4 antibody
        • Cytotoxic T lymphocyte antigen 4 antibody
        • Cytotoxic T lymphocyte associated 4 antibody
        • Cytotoxic T lymphocyte associated 4, soluble isoform, included antibody
        • Cytotoxic T lymphocyte associated antigen 4 antibody
        • Cytotoxic T lymphocyte associated antigen 4 short spliced form antibody
        • Cytotoxic T lymphocyte associated protein 4 antibody
        • Cytotoxic T lymphocyte associated serine esterase 4 antibody
        • Cytotoxic T lymphocyte protein 4 antibody
        • Cytotoxic T-lymphocyte protein 4 antibody
        • Cytotoxic T-lymphocyte-associated antigen 4 antibody
        • GRD4 antibody
        • GSE antibody
        • ICOS antibody
        • IDDM12 antibody
        • insulin-dependent diabetes mellitus 12 antibody
        • Ligand and transmembrane spliced cytotoxic T lymphocyte associated antigen 4 antibody
        • OTTHUMP00000216623 antibody
        see all

      Images

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTLA4 antibody [CAL49] (ab237712)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTLA4 antibody [CAL49] (ab237712)

        Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CTLA4 with ab237712 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining on human tonsil is observed. Counter stained with hematoxylin.

        Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

        Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins.

        The section was incubated with ab237712 for 30 mins at room temperature.

        The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

      • Western blot - Anti-CTLA4 antibody [CAL49] (ab237712)
        Western blot - Anti-CTLA4 antibody [CAL49] (ab237712)
        All lanes : Anti-CTLA4 antibody [CAL49] (ab237712) at 1/1000 dilution

        Lane 1 : Untreated human PBMC (human peripheral blood mononuclear cell) whole cell lysate
        Lane 2 : Human PBMC (treated with 10µg/ml PHA for 2 days) whole cell lysate
        Lane 3 : Untreated mouse splenocyte whole cell lysate
        Lane 4 : Mouse splenocyte (treated with 2.5µg/ml Concanavalin A (ConA) for 3 days) whole cell lysate

        Lysates/proteins at 20 µg per lane.

        Secondary
        All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

        Predicted band size: 25 kDa
        Observed band size: 25 kDa


        Exposure time: 3 minutes


        Blocking and dilution buffer: 5% NFDM/TBST.

      • Flow Cytometry - Anti-CTLA4 antibody [CAL49] (ab237712)
        Flow Cytometry - Anti-CTLA4 antibody [CAL49] (ab237712)

        Flow cytometric analysis of 2% paraformaldehyde-fixed, 0.1% Tween 20 permeabilized mouse splenocytes (treated with 2.5μg/ml Concanavalin A (ConA) for 3 days) cells labeling CTLA4 with ab237712 at 1/400 (Right) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (Left).

        Cells were surface stained with anti-CD3 conjugated to Alexa Fluor® 647. Then fixed with 2% PFA for 10min followed by intracellular staining with rabbit IgG (Left) and ab237712 (Right).

        Goat Anti-Rabbit IgG Fc (Alexa Fluor® 488) preadsorbed (ab150097), at 1/2000 dilution was used as the secondary antibody.

      • Immunoprecipitation - Anti-CTLA4 antibody [CAL49] (ab237712)
        Immunoprecipitation - Anti-CTLA4 antibody [CAL49] (ab237712)

        CTLA4 was immunoprecipitated from 0.35 mh human tonsil lysate with ab237712 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab237712 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as secondary antibody at 1/1000 dilution.

        Lane 1: Human tonsil lysate 10 μg (Input).
        Lane 2: ab237712 IP in human tonsil lysate.
        Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab237712 in human tonsil lysate.

        Blocking and dilution buffer and concentration: 5% NFDM/TBST.
        Exposure time: 30 seconds.

      • Flow Cytometry - Anti-CTLA4 antibody [CAL49] (ab237712)
        Flow Cytometry - Anti-CTLA4 antibody [CAL49] (ab237712)

        Flow cytometric analysis of 2% paraformaldehyde-fixed, 0.1% Tween 20 permeabilized human PBMC (peripheral blood mononuclear cell) (treated with 10μg/ml PHA for 2 days) cells labeling CTLA4 with ab237712 at 1/400 (Right) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (Left).

        Cells were surface stained with anti-CD3 conjugated to Alexa Fluor® 647. Then fixed with 2% PFA for 10min followed by intracellular staining with rabbit IgG (Left) and ab237712 (Right).

        Goat Anti-Rabbit IgG Fc (Alexa Fluor® 488) preadsorbed (ab150097), at 1/2000 dilution was used as the secondary antibody.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTLA4 antibody [CAL49] (ab237712)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTLA4 antibody [CAL49] (ab237712)

        Formalin-fixed, paraffin-embedded human breast carcinoma tissue stained for CTLA4 using ab237712 at 0.25 μg/ml in immunohistochemical analysis.

        Incubate with primary antibody for 75 minutes at room temperature.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTLA4 antibody [CAL49] (ab237712)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTLA4 antibody [CAL49] (ab237712)

        Formalin-fixed, paraffin-embedded human lymph node tissue stained for CTLA4 using ab237712 at 0.25 μg/ml in immunohistochemical analysis.

        Incubate with primary antibody for 75 minutes at room temperature.

      Protocols

      To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

      Click here to view the general protocols

      References

      Publishing research using ab237712? Please let us know so that we can cite the reference in this datasheet.

      ab237712 has not yet been referenced specifically in any publications.

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