Anti-TIM 3 antibody [EPR22241]

9 product images

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIM 3 antibody [EPR22241] (ab241332)

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue labeling TIM 3 with ab241332 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic and membranous staining on both infiltrated immunocytes and tumor cells of human lung cancer (PMID: 22586058) 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 using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIM 3 antibody [EPR22241] (ab241332)

  Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling TIM 3 with ab241332 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic and membranous staining on subsets of immune cells of mouse spleen 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 using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).

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  Immunoprecipitation - Anti-TIM 3 antibody [EPR22241] (ab241332)

  TIM 3 was immunoprecipitated from 0.35 mg of Daudi (human Burkitt's lymphoma cell line) whole cell lysate with ab241332 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab241332 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/1000 dilution.
  

  Lane 1: Daudi lysate 10 μg (Input).
  Lane 2: ab241332 IP in Daudi whole cell lysate (+).
  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab241332 in Daudi whole cell lysate (-).

  Blocking and dilution buffer and concentration: 5% NFDM/TBST.
  Exposure time: 3 minutes.

  All lanes: Immunoprecipitation - Anti-TIM 3 antibody [EPR22241] (ab241332)

  Predicted band size: 33 kDa

  Observed band size: 50 kDa

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIM 3 antibody [EPR22241] (ab241332)

  Immunohistochemical analysis of paraffin-embedded human liver tissue labeling TIM 3 with ab241332 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic and membranous staining on Kupffer cells of human liver (PMID: 27192565) 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 using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIM 3 antibody [EPR22241] (ab241332)

  Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling TIM 3 with ab241332 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic and membranous staining on subsets of immune cells of rat spleen 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 using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).

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  Western blot - Anti-TIM 3 antibody [EPR22241] (ab241332)

  Blocking and dilution buffer: 5% NFDM/TBST.
  

  Lanes 3 & 4 were developed using a higher sensitivity ECL substrate.

  All lanes: Western blot - Anti-TIM 3 antibody [EPR22241] (ab241332) at 1/1000 dilution

  Lane 1: Daudi (human Burkitt's lymphoma cell line) whole cell lysate at 20 µg

  Lane 2: Daudi whole cell lysate (treated with PNGase F) at 20 mg/mL

  Lane 3: RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 20 µg

  Lane 4: RAW 264.7 whole cell lysate (treated with PNGase F) at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  Developed using the ECL technique.

  Predicted band size: 33 kDa

  Observed band size: 40 kDa, 50-60 kDa

  Exposure time: 70s

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIM 3 antibody [EPR22241] (ab241332)

  Tissue Microarrays stained for "Anti-TIM 3 antibody [EPR22241]" using "ab241332"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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). The sections were incubated with ab241332 at +4°C overnight followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIM 3 antibody [EPR22241] (ab241332)

  Tissue Microarrays stained for "Anti-TIM 3 antibody [EPR22241]" using "ab241332"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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). The sections were incubated with ab241332 at +4°C overnight followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

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  Multiplex immunohistochemistry - Anti-TIM 3 antibody [EPR22241] (ab241332)

  This data was developed using Anti-TIM 3 antibody [EPR22241] - BSA and Azide free ab242080, the same antibody clone in a different buffer formulation.
  
  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human tonsil labelling PD1 with Anti-PD1 antibody [EPR23119-111] ab243644 at 1/500 dilution (1.02 µg/mL) (D), CD8 with Anti-CD8 alpha antibody [SP239] ab178089 at 1/100 dilution (0.83 μg/ml) (C) and TIM 3 with Anti-TIM 3 antibody [EPR22241] - BSA and Azide free ab242080 at 1/500 dilution (2.09 μg/ml) (B). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.  
  
  Panel A: merged staining of anti-TIM 3 (green; Opal™690), anti-CD8 (red; Opal™520) and anti-PD1 (gray; Opal™570) on human tonsil.
  Panel B: anti-TIM 3 stained on membrane of a subset of immune cells.
  Panel C: anti-CD8 stained on membrane of a subset of T cells.
  Panel D: anti-PD1 stained on membrane of a subset of lymphocytes.
  
  The section was incubated in three rounds of staining: in the order of Anti-TIM 3 antibody [EPR22241] - BSA and Azide free ab242080, Anti-PD1 antibody [EPR23119-111] ab243644 and Anti-CD8 alpha antibody [SP239] ab178089 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.
  
  The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.