Anti-Androgen Receptor antibody [EP670Y]

• WB

Lab

Western blot - Anti-Androgen Receptor antibody [EP670Y] (AB52615)

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

This antibody works better in 1%SDS Hot Lysates in WB. For Lysate preparation protocol, please refer to the protocol section of the website and/or here.

All lanes:

Western blot - Anti-Androgen Receptor antibody [EP670Y] (ab52615) at 1/5000 dilution

Lane 1:

22Rv1 (Human prostate carcinoma epithelial cell) whole cell lysates prepared in 1%SDS Hot lysis method at 15 µg

Lane 2:

22Rv1 (Human prostate carcinoma epithelial cell) whole cell lysates prepared in RIPA lysis method at 15 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: 99 kDa

Observed band size: 110 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-Androgen Receptor antibody [EP670Y] (AB52615)

Lanes 1 - 2 : Merged signal (red and green). Green - ab52615 observed at 120 kDa. Red - loading control, ab8245, observed at 37 kDa.

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab52615 and ab8245 (loading control) overnight at 4°C. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at a 1 : 10000 dilution for 1hr at room temperature and then imaged.

All lanes:

Western blot - Anti-Androgen Receptor antibody [EP670Y] (ab52615) at 1/1000 dilution

Lane 1:

LnCAP whole cell lysate (positive control) at 20 µg

Lane 2:

PC3 whole cell lysate (negative control) at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 99 kDa

Observed band size: 120 kDa,58 kDa

false

• WB

Lab

Western blot - Anti-Androgen Receptor antibody [EP670Y] (AB52615)

Blocking/Diluting buffer : 5% NFDM/TBST

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

All lanes:

Western blot - Anti-Androgen Receptor antibody [EP670Y] (ab52615) at 1/1000 dilution

Lane 1:

Mouse testis lysate at 20 µg

Lane 2:

Rat testis lysate at 20 µg

Lane 3:

Mouse liver lysate at 20 µg

Lane 4:

Rat liver 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: 99 kDa

Observed band size: 110 kDa

false

Exposure time: 1min

• WB

Lab

Western blot - Anti-Androgen Receptor antibody [EP670Y] (AB52615)

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Androgen Receptor antibody [EP670Y] (ab52615) at 1/2000 dilution

Lane 1:

Mouse testis tissue lysate at 10 µg

Lane 2:

Rat testis tissue lysate at 10 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 1/50000 dilution

Predicted band size: 99 kDa

Observed band size: 110 kDa

false

• WB

Unknown

Western blot - Anti-Androgen Receptor antibody [EP670Y] (AB52615)

All lanes:

Western blot - Anti-Androgen Receptor antibody [EP670Y] (ab52615) at 1/10000 dilution

All lanes:

LnCaP cell lysate at 10 µg

Secondary

All lanes:

HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

Predicted band size: 99 kDa

Observed band size: 99 kDa

false

• WB

CiteAb

Western blot - Anti-Androgen Receptor antibody [EP670Y] (AB52615)

Androgen Receptor western blot using anti-Androgen Receptor antibody [EP670Y] ab52615. Publication image and figure legend from Tieszen, C. R., Goyeneche, A. A., et al., 2011, BMC Cancer, PubMed 21619605.

ab52615 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab52615 please see the product overview.

Expression of progesterone, androgen, glucocorticoid and estrogen receptors in cancer cells exposed to MF. Cells were exposed to either vehicle or the IC50 concentration of MF specific to each cell line for 24 h. Cells were subsequently harvested, lysed, and whole-protein extracts (50 μg for GR and AR; 100 μg for PR and ER-α) were separated by electrophoresis. Immunoblots were then probed with the indicated antibodies (for PR : †clone hPRa7, Thermo Fisher Scientific; ‡ #1483-1, Epitomics); two commercially available antibodies for human PR were used to strengthen the reliability of the results. β-actin was included as a control for protein loading. Because AR, GR-α and GR-β were immunoblotted in one membrane and PR-A, PR-B and ER-α were blotted in a separate membrane, each membrane was blotted separately with anti-β-actin. This experiment was repeated twice with similar outcome. MDA-231 means MDA-MB-231.

false