All lanes : Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273) at 1/1000 dilution

Lane 1 : Mouse testis lysate
Lane 2 : Rat testis lysate
Lane 3 : Mouse liver lysate
Lane 4 : Rat liver lysate

Lysates/proteins at 20 µg per lane.

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

Predicted band size: 98 kDa
Observed band size: 110 kDa why is the actual band size different from the predicted?


Exposure time: 20 seconds

Blocking/Diluting buffer: 5% NFDM/TBST

Loading Control: Rabbit monoclonal [EPR16891] to GAPDH (ab181602)

Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273) at 1/5000 dilution (purified) + Mouse prostate lysates at 15 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 98 kDa

Blocking and diluting buffer: 5% NFDM/TBST

Texas red (Tx-Red) indirect immunofluorescent detection of Androgen Receptor using ab133273 in (A) EP156T-AR, (B) 957E/hTERT-AR and (C) LNCaP cells. The cells were treated with ± 1 nM R1881 for 24 hours.

Formalin-fixed, paraffin-embedded tissues labeling Androgen Receptor with ab133273. Left: Tumor from orthotopic injectionof PTEN KD/TE cells. Right: Human postate cancer tissue.

From Figure 3 of Shao et al/

Shao et al Oncotarget. 2018 Mar 6; 9(18): 14456–14471.Published online 2018 Feb 12. doi: 10.18632/oncotarget.24470

Reproduced under the terms of the Creative Commons Attribution License 3.0 (CC BY 3.0).

Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling Androgen receptor with purified ab133273 at 1:100 dilution (1.3μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1:200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor^® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

Immunohistochemical analysis of paraffin-embedded human prostatic adenocarcinoma tissue labelling Androgen Receptor  using  unpurified ab133273 at 1/100 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

All lanes : Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273) at 1/20000 dilution

Lane 1 : LNCaP (Human prostate carcinoma epithelial cell) whole cell lysates prepared in RIPA lysis method
Lane 2 : LNCaP (Human prostate carcinoma epithelial cell) whole cell lysates prepared in 1%SDS Hot lysis method
Lane 3 : 22Rv1 (Human prostate carcinoma epithelial cell) whole cell lysates prepared in RIPA lysis method
Lane 4 : 22Rv1 (Human prostate carcinoma epithelial cell) whole cell lysates prepared in 1%SDS Hot lysis method

Lysates/proteins at 1/15 dilution per lane.

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

Predicted band size: 98 kDa
Observed band size: 120 kDa why is the actual band size different from the predicted?


Exposure time: 10 seconds

Blocking/Diluting buffer - 5% NFDM/TBST

The androgen receptor variant band detected in 22RV1 cells is reported by PMID: 22315407.

We recommend you to try both RIPA and 1%SDS Hot lysis preparation methods to get desired bands.

Immunofluorescent staining of LnCAP cells labelling Androgen Receptor using unpurified ab133273, at 1/100 dilution

All lanes : Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273) at 1/5000 dilution (purified)

Lane 1 : LNCaP (Human prostate carcinoma epithelial cell) whole cell lysates
Lane 2 : Rat prostate lysates

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

Predicted band size: 98 kDa

Blocking and diluting buffer: 5% NFDM/TBST

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat testis tissue sections labeling Androgen Receptor with Purified ab133273 at 1:500 dilution (0.25 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse testis tissue sections labeling Androgen Receptor with Purified ab133273 at 1:500 dilution (0.25 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostatic hyperplasia tissue sections labeling Androgen Receptor with Purified ab133273 at 1:500 dilution (0.25 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273) at 1/1000 dilution (unpurified) + LNCaP (Human prostate carcinoma epithelial cell) at 10 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 98 kDa
Observed band size: 110 kDa why is the actual band size different from the predicted?


Exposure time: 3 seconds

Blocking/Diluting buffer 5% NFDM/TBST

Immunohistochemical analysis of paraffin-embedded human prostate tissue labelling Androgen Receptor using unpurified ab133273, at1/100 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Unpurified  ab133273 showing positive staining in Breast carcinoma tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Unpurified  ab133273 showing negative staining in Normal brain tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Unpurified  ab133273 showing negative staining in Normal tonsil tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Unpurified ab133273 showing positive staining in Endometrial carcinoma tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Unpurified ab133273 showing negative staining in Normal breast tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Unpurified ab133273 showing negative staining in Normal colon tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Unpurified ab133273 showing negative staining in Ovarian carcinoma tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Unpurified ab133273 showing negative staining in Colonic adenocarcinoma tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

 Unpurified ab133273 showing negative staining in Normal liver tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Unpurified  ab133273 showing positive staining in Prostatic carcinoma T3 tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.