Anti-TIM 3 antibody [EPR22241]

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-TIM 3 antibody [EPR22241] (AB241332)

This data was developed using ab242080, the same antibody clone in a different buffer formulation. Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human tonsil labelling PD1 with ab243644 at 1/500 dilution (1.02 µg/mL) (D), CD8 with ab178089 at 1/100 dilution (0.83 μg/ml) (C) and TIM 3 with 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 ab242080, ab243644 and 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.

• IHC-P

Supplier Data

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 ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

Supplier Data

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 ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

Lab

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 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).

• IP

Supplier Data

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) (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 (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

false

• IHC-P

Supplier Data

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 ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

Lab

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 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).

• IHC-P

Supplier Data

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 ab93684 (Tris/EDTA buffer, pH 9.0).

• WB

Supplier Data

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) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 33 kDa

Observed band size: 40 kDa,50-60 kDa

true

Exposure time: 70s