Anti-Fas antibody [EPR24898-74]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fas antibody [EPR24898-74] (AB271016)

Immunohistochemical analysis of paraffin-embedded sections of mouse thymus labelling Fas with ab271016 at 1/100 dilution followed by ready to use secondary antibody LeicaDS9800 (Bond™ Polymer Refine Detection). Membranous staining on mouse thymus. Counterstained with Hematoxylin.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins was used. The section was incubated with ab271016 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

Secondary antibody only control : PBS instead of primary antibody followed by LeicaDS9800 (Bond™ Polymer Refine Detection) as a secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fas antibody [EPR24898-74] (AB271016)

Immunohistochemical analysis of paraffin-embedded sections of mouse spleen labelling Fas with ab271016 at 1/100 dilution followed by ready to use secondary antibody LeicaDS9800 (Bond™ Polymer Refine Detection). Membranous staining on mouse spleen. Counterstained with Hematoxylin.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins was used. The section was incubated with ab271016 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

Secondary antibody only control : PBS instead of primary antibody followed by LeicaDS9800 (Bond™ Polymer Refine Detection) as a secondary antibody.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Fas antibody [EPR24898-74] (AB271016)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse thymus (fresh) tissue labelling Fas with ab 271016 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150081) at 1/1000 dilution (Green). Positive staining on mouse thymus. The nuclear counter stain was DAPI (Blue).

Secondary antibody only control : Used PBS instead of primary antibody follwed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150081) at 1/1000 dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Fas antibody [EPR24898-74] (AB271016)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse liver (fresh) tissue labelling Fas with ab 271016 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150081) at 1/1000 dilution (Green). Positive staining on mouse liver. The nuclear counter stain was DAPI (Blue).

Secondary antibody only control : Used PBS instead of primary antibody followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150081) at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fas antibody [EPR24898-74] (AB271016)

Immunohistochemical analysis of paraffin-embedded sections of mouse lung cancer labelling Fas with ab271016 at 1/100 dilution followed by ready to use secondary antibody LeicaDS9800 (Bond™ Polymer Refine Detection). Membranous staining on mouse lung cancer tissue. Counterstained with Hematoxylin.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins was used. The section was incubated with ab271016 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

Secondary antibody only control : PBS instead of primary antibody followed by LeicaDS9800 (Bond™ Polymer Refine Detection) as a secondary antibody.

• IP

Supplier Data

Immunoprecipitation - Anti-Fas antibody [EPR24898-74] (AB271016)

Fas was immunoprecipitated from mouse thymus tissue lysate with ab271016 at 1/30 dilution (2 μg in 0.35 mg lysates). Western blot was performed from innunoprecipitate using ab271016 at 1/1000 dilution. Secondary antibody VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/5000 dilution.

Lane 1 : Mouse thymus tissue lysate (Input) 10 μg

Lane 2 : ab271016 IP in Mouse thymus tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab271016 in mouse thymus tissue lysate

Exposure time : 5.5 seconds

The 20KDa band could be a degradation or cleavage product, as demonstrated by the use of fresh lysate in the WB data.

All lanes:

Immunoprecipitation - Anti-Fas antibody [EPR24898-74] (ab271016)

Predicted band size: 37 kDa

Observed band size: 45 kDa

false

• WB

Supplier Data

Western blot - Anti-Fas antibody [EPR24898-74] (AB271016)

5% NFDM/TBST was used as a blocking and diluting buffer.

Exposure times :

Lane 1-2 : 37 seconds;
Lane 3 : 81 seconds.

Lanes 2-3 lysate was made fresh and used immediately to minimize protein degradation.

The 20 kDa band in lane 1 could be a degradation or cleavage product.

All lanes:

Western blot - Anti-Fas antibody [EPR24898-74] (ab271016) at 1/1000 dilution

Lane 1:

Mouse thymus tissue lysate at 20 µg

Lane 2:

Mouse thymus tissue fresh lysate at 20 µg

Lane 3:

A20 (mouse reticulum sarcoma B lymphocyte) whole cell fresh lysate 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 1/100000 dilution

Predicted band size: 37 kDa

Observed band size: 45 kDa

false

• WB

Supplier Data

Western blot - Anti-Fas antibody [EPR24898-74] (AB271016)

5% NFDM/TBST was used as a blocking and diluting buffer.

Exposure times :

Lane 1, 3 : 26 seconds;
Lane 2 : 136 seconds.

The observed MW are consistent with what has been described in the literature (PMID : 28883393).

All lanes:

Western blot - Anti-Fas antibody [EPR24898-74] (ab271016) at 1/1000 dilution

Lane 1:

Mouse liver tissue lysate at 20 µg

Lane 2:

Mouse heart tissue lysate at 20 µg

Lane 3:

Mouse lung tissue lysate 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: 37 kDa

Observed band size: 45 kDa

false

• WB

Supplier Data

Western blot - Anti-Fas antibody [EPR24898-74] (AB271016)

5% NFDM/TBST was used as blocking and diluting buffer.

The observed MW are consistent with what has been described in the literature (PMID : 28883393).

Low expression : stomach, skin (PMID : 31582729; PMID : 11106570).

All lanes:

Western blot - Anti-Fas antibody [EPR24898-74] (ab271016) at 1/1000 dilution

Lane 1:

Mouse spleen tissue lysate at 20 µg

Lane 2:

Mouse stomach tissue lysate at 20 µg

Lane 3:

Mouse skin tissue lysate 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 1/100000 dilution

Predicted band size: 37 kDa

Observed band size: 45 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-Fas antibody [EPR24898-74] (AB271016)

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Fas antibody [EPR24898-74] (ab271016) at 1/1000 dilution

Lane 1:

Mouse pancreas tissue lysate at 20 µg

Lane 2:

Mouse placenta tissue lysate at 20 µg

Lane 3:

Mouse uterus tissue lysate at 20 µg

Lane 4:

J774A.1 (mouse reticulum cell sarcoma monocyte macrophage) whole cell lysate at 20 µg

Lane 5:

L929 (mouse connective tissue fibroblast) whole cell lysate 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: 37 kDa

Observed band size: 45 kDa

false

Exposure time: 180s