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anti-CTLA4 antibody [CAL49] ab251599 is a carrier free rabbit monoclonal antibody that is used in CTLA4 western blotting, IHC, flow cytometry and Immunoprecipitation. Suitable for human, monkey and mouse samples.

- PBS only, conjugation-ready, removing anything extra from your antibodies, greater flexibility in assay design
- Validated on the Leica BOND™ RX automated IHC staining platform for CTLA4 IHC
- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Antibody clone CAL49 is cited in over 30 publications


Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTLA4 antibody [CAL49] - BSA and Azide free (AB251599), expandable thumbnail
  • Immunoprecipitation - Anti-CTLA4 antibody [CAL49] - BSA and Azide free (AB251599), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-CTLA4 antibody [CAL49] - BSA and Azide free (AB251599), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-CTLA4 antibody [CAL49] - BSA and Azide free (AB251599), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTLA4 antibody [CAL49] - BSA and Azide free (AB251599), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Constituents: PBS

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPWBICC/IFFlow Cyt (Intra)IHC-P
Human
Tested
Tested
Not recommended
Tested
Tested
Mouse
Expected
Tested
Not recommended
Tested
Expected
Rhesus monkey
Expected
Expected
Not recommended
Expected
Tested

Tested
Tested

Species

Human

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species

Mouse, Rhesus monkey

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse, Human

Dilution info

-

Notes

-

Expected
Expected

Species

Rhesus monkey

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Mouse, Human, Rhesus monkey

Dilution info

-

Notes

-

Tested
Tested

Species

Mouse, Human

Dilution info

-

Notes

-

Expected
Expected

Species

Rhesus monkey

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species

Rhesus monkey

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species

Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Associated Products

Select an associated product type

1 product for Alternative Version

5 products for Alternative Product

Target data

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.

Alternative names

CD152, CTLA4, CD152, Cytotoxic T-lymphocyte protein 4, Cytotoxic T-lymphocyte-associated antigen 4, CTLA-4

Recommended products

anti-CTLA4 antibody [CAL49] ab251599 is a carrier free rabbit monoclonal antibody that is used in CTLA4 western blotting, IHC, flow cytometry and Immunoprecipitation. Suitable for human, monkey and mouse samples.

- PBS only, conjugation-ready, removing anything extra from your antibodies, greater flexibility in assay design
- Validated on the Leica BOND™ RX automated IHC staining platform for CTLA4 IHC
- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Antibody clone CAL49 is cited in over 30 publications

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free

Yes

Clone number

CAL49

Purification technique

Affinity purification Protein A

Concentration
Loading...
Purification notes

Purity is greater than 99%.

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

Notes

ab251599 is the carrier-free version of Anti-CTLA4 antibody [CAL49] ab237712.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

CTLA-4 also known as CD152 is an immune checkpoint receptor with a molecular weight of approximately 34 kDa. This protein is expressed on the surface of T-cells especially after activation and sometimes on regulatory T-cells (Tregs). CTLA-4 competes with CD28 for binding to the same ligands CD80 and CD86 on antigen-presenting cells. Unlike CD28 which stimulates T-cell activation CTLA-4 sends inhibitory signals to downregulate immune responses. By doing this CTLA-4 helps maintain immune system homeostasis and prevents autoimmune reactions.

Biological function summary

The CTLA-4 protein functions as a critical regulator of the immune system. It interacts with CD80/CD86 in a complex that effectively transmits inhibitory signals to T-cells. CTLA-4 controls the amplitude of the initial activation of T-cells ensuring that the body's immune responses are kept under control. In regulatory T-cells CTLA-4 engagement contributes to their immunosuppressive functions through which they maintain tolerance to self-antigens and prevent overactive immune reactions.

Pathways

CTLA-4 involves itself in the adaptive immune response pathways specifically the regulation of T-cell activity. CTLA-4's interaction with CD80/CD86 modulates pathways associated with TCR (T-cell receptor) signaling. When CTLA-4 binds its ligands it recruits phosphatases such as SHP-2 that dephosphorylate signaling proteins leading to the downregulation of T-cell interaction. This pathway interaction exemplifies how CTLA-4 acts in concert with proteins like CD28 to finely tune immune responses.

Associated diseases and disorders

CTLA-4 plays a role in autoimmune diseases and cancer. The dysregulation of CTLA-4 function can lead to autoimmune disorders where the immune system attacks the body's own tissue. In cancer tumor cells may manipulate CTLA-4 pathways to evade immune surveillance. Targeted therapies like anti-CTLA-4 antibodies such as ipilimumab have been developed to block CTLA-4 aiming to enhance the immune system’s ability to attack cancer cells. These treatments highlight the role of CTLA-4 in balancing immune activity and the potential to alter this balance for therapeutic benefit.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

9 product images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTLA4 antibody [CAL49] - BSA and Azide free (ab251599), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTLA4 antibody [CAL49] - BSA and Azide free (ab251599)

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CTLA4 with Anti-CTLA4 antibody [CAL49] 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 Anti-CTLA4 antibody [CAL49] ab237712 for 30 mins at room temperature.

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (Anti-CTLA4 antibody [CAL49] ab237712).

  • Immunoprecipitation - Anti-CTLA4 antibody [CAL49] - BSA and Azide free (ab251599), expandable thumbnail

    Immunoprecipitation - Anti-CTLA4 antibody [CAL49] - BSA and Azide free (ab251599)

    CTLA4 was immunoprecipitated from 0.35 mh human tonsil lysate with Anti-CTLA4 antibody [CAL49] ab237712 at 1/30 dilution. Western blot was performed from the immunoprecipitate using Anti-CTLA4 antibody [CAL49] ab237712 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (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: Anti-CTLA4 antibody [CAL49] ab237712 IP in human tonsil lysate.
    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-CTLA4 antibody [CAL49] ab237712 in human tonsil lysate.

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (Anti-CTLA4 antibody [CAL49] ab237712).

    All lanes: Immunoprecipitation - Anti-CTLA4 antibody [CAL49] (Anti-CTLA4 antibody [CAL49] ab237712)

    Predicted band size: 24 kDa

    Observed band size: 25 kDa

  • Flow Cytometry (Intracellular) - Anti-CTLA4 antibody [CAL49] - BSA and Azide free (ab251599), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-CTLA4 antibody [CAL49] - BSA and Azide free (ab251599)

    Intracellular flow cytometric analysis of2% paraformaldehyde-fixed, 0.1% Tween 20 permeabilized mouse splenocytes (treated with 2.5μg/ml Concanavalin A (ConA) for 3 days) cells labeling CTLA4 with Anti-CTLA4 antibody [CAL49] ab237712 at 1/400 (Right) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (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 Anti-CTLA4 antibody [CAL49] ab237712 (Right).

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (Anti-CTLA4 antibody [CAL49] ab237712).

  • Flow Cytometry (Intracellular) - Anti-CTLA4 antibody [CAL49] - BSA and Azide free (ab251599), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-CTLA4 antibody [CAL49] - BSA and Azide free (ab251599)

    Intracellular 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 Anti-CTLA4 antibody [CAL49] ab237712 at 1/400 (Right) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (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 Anti-CTLA4 antibody [CAL49] ab237712 (Right).

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (Anti-CTLA4 antibody [CAL49] ab237712).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTLA4 antibody [CAL49] - BSA and Azide free (ab251599), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTLA4 antibody [CAL49] - BSA and Azide free (ab251599)

    Formalin-fixed, paraffin-embedded human breast carcinoma tissue stained for CTLA4 using Anti-CTLA4 antibody [CAL49] ab237712 at 0.25 μg/ml in immunohistochemical analysis.

    Incubate with primary antibody for 75 minutes at room temperature.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (Anti-CTLA4 antibody [CAL49] ab237712).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTLA4 antibody [CAL49] - BSA and Azide free (ab251599), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTLA4 antibody [CAL49] - BSA and Azide free (ab251599)

    Formalin-fixed, paraffin-embedded human lymph node tissue stained for CTLA4 using Anti-CTLA4 antibody [CAL49] ab237712 at 0.25 μg/ml in immunohistochemical analysis.

    Incubate with primary antibody for 75 minutes at room temperature.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (Anti-CTLA4 antibody [CAL49] ab237712).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTLA4 antibody [CAL49] - BSA and Azide free (ab251599), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTLA4 antibody [CAL49] - BSA and Azide free (ab251599)

    Tissue Microarrays stained for "Anti-CTLA4 antibody [CAL49]" using "Anti-CTLA4 antibody [CAL49] ab237712"in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 30 minutes. The sections were incubated with Anti-CTLA4 antibody [CAL49] ab237712 for 30 mins at room temperature followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTLA4 antibody [CAL49] - BSA and Azide free (ab251599), expandable thumbnail
    This image is courtesy of Dr. Chi Ngai Chan

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTLA4 antibody [CAL49] - BSA and Azide free (ab251599)

    Immunohistochemical analysis of paraffin-embeded Rhesus monkey tonsil tissue labeling CTLA4 with ab251599 followed by Polink 1 Polymer HRP anti-Rabbit IgG.

    Heat mediated antigen retrieval-Buffer/Enzyme Used: Dako pH9.

  • Flow Cytometry (Intracellular) - Anti-CTLA4 antibody [CAL49] - BSA and Azide free (ab251599), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-CTLA4 antibody [CAL49] - BSA and Azide free (ab251599)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CTLA4 antibody [CAL49] ab237712).

    Flow cytometry staining of human peripheral blood mononuclear cells (PBMCs) (top) or PBMCs treated with PHA, 48 Hours, 5 µg/mL (bottom), with Anti-CTLA4 antibody [CAL49] ab237712 (right) or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (left). Cells were fixed and permeabilised with BD Cytofix/Cytoperm™ for 20 min. PBMCs were incubated for 30 min on ice in 1x PBS containing 10 µg/ml human IgG and 10 % normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody Anti-CTLA4 antibody [CAL49] ab237712 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 106 in 100 µl at 0.2 μg/ml (1/10,300)) for 30 min at 22°C . The cells were simultaneously stained with CD3.

    The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/2000 for 30min at 22°C

    Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on live cells.

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