Anti-CTLA4 antibody [CAL49]

9 product images

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  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.

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  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) (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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab237712 in human tonsil lysate.

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

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

  Predicted band size: 24 kDa

  Observed band size: 25 kDa

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  Western blot - Anti-CTLA4 antibody [CAL49] (ab237712)

  Blocking and dilution buffer: 5% NFDM/TBST.

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

  Lane 1: Untreated human PBMC (human peripheral blood mononuclear cell) whole cell lysate at 20 µg

  Lane 2: Human PBMC (treated with 10μg/ml PHA for 2 days) whole cell lysate at 20 µg

  Lane 3: Untreated mouse splenocyte whole cell lysate at 20 µg

  Lane 4: Mouse splenocyte (treated with 2.5μg/ml Concanavalin A (ConA) for 3 days) whole cell lysate at 20 µg

  Secondary

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

  Predicted band size: 24 kDa

  Observed band size: 25 kDa

  Exposure time: 3min

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

  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 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 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.

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

  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 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 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.

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  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.

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  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.

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  This image is courtesy of Dr. Chi Ngai Chan

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

  Immunohistochemical analysis of paraffin-embeded Rhesus monkey tonsil tissue labeling CTLA4 with Anti-CTLA4 antibody [CAL49] - BSA and Azide free ab251599 followed by Polink 1 Polymer HRP anti-Rabbit IgG.

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

  This data was developed using the same antibody clone in a different buffer formulation (Anti-CTLA4 antibody [CAL49] - BSA and Azide free ab251599).

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

  Tissue Microarrays stained for "Anti-CTLA4 antibody [CAL49]" using "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 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.