Recombinant 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. 

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.  

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.

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. 

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).

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).

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. 

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.

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

Exposure times:

Lane 1, 3: 92 seconds;
Lane 2 : 10 seconds. 

Low expression: L929 (PMID: 7523113).

The 20 kDa band in lane 1-2 could be a degradation or cleavage product.

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.