All lanes : Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] (ab275745) at 1/1000 dilution

Lane 1 : Human testis tissue lysate
Lane 2 : Human brain tissue lysate
Lane 3 : Human skeletal muscle tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/1000 dilution

Predicted band size: 51 kDa
Observed band size: 52 kDa why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Low expression: human brain, human skeletal muscle (Database:GTExPortal, HPA).

Exposure time: 103 seconds.

 

All lanes : Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] (ab275745) at 1/1000 dilution

Lane 1 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lane 2 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution

Predicted band size: 51 kDa
Observed band size: 52 kDa why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Exposure time: 70 seconds.

 

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling Retinoic Acid Receptor alpha with ab275745 at 1/1000 (2.71 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nuclear staining on sertoli cells of human testis. The section was incubated with ab275745 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

Retinoic Acid Receptor alpha was immunoprecipitated from 0.35 mg HEK-293 (human embryonic kidney epithelial cell) whole cell lysate with ab275745 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab275745 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: HEK-293 (human embryonic kidney epithelial cell) whole cell lysate 10 ug

Lane 2: ab275745 IP in HEK-293 whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab275745 in HEK-293 whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 84 seconds.

 

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling Retinoic Acid Receptor alpha with ab275745 at 1/1000 (0.542 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nuclear staining on sertoli cells of rat testis. The section was incubated with ab275745 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

All lanes : Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] (ab275745) at 1/1000 dilution

Lane 1 : MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : U937 (human histiocytic lymphoma monocyte) whole cell lysate
Lane 3 : HEK-293 (human embryonic kidney epithelial cell) whole cell lysate
Lane 4 : MOLT-4 (human lymphoblastic leukemia T lymphoblast) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution

Predicted band size: 51 kDa
Observed band size: 52 kDa why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Low expression: MOLT-4.(Database:HPA).

Exposure time: Lane 1: 8 seconds Lane2-4: 70 seconds.

 

Retinoic Acid Receptor alpha was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate with ab275745 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab275745 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 ug

Lane 2: ab275745 IP in NIH/3T3 whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab275745 in NIH/3T3 whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 84 seconds.

 

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling Retinoic Acid Receptor alpha with ab275745 at 1/1000 (0.542 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nuclear staining on sertoli cells of mouse testis (PMID:16210368). The section was incubated with ab275745 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).