Anti-P Glycoprotein antibody [EPR10364-57] (ab170904)

Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: P Glycoprotein knockout HAP1 whole cell lysate (20 µg)
Lane 3: HepG2 whole cell lysate (20 µg)

Lanes 1 - 3: Merged signal (red and green). Green - ab170904 observed at 160 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab170904 was shown to specifically react with P Glycoprotein when P Glycoprotein knockout samples were used. Wild-type and P Glycoprotein knockout samples were subjected to SDS-PAGE. ab170904 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10000 dilution respectively.

Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

Blocking and diluting buffer: 5% NFDM/TBST.

Immunohistochemical staining of paraffin embedded human kidney with purified ab170904 at a working dilution of 1/100. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. Counterstained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.

PBS was used instead of the primary antibody as the negative control (inset).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue sections labeling P Glycoprotein with purified ab170904 at 1:1200 dilution (0.24 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0) ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Hematoxylinwas used as a counterstain.

Negative control: PBS instead of the primary antibody (inset).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human hepatocellular carcinoma tissue labeling P Glycoprotein with unpurified ab170904 at 1/250 dilution.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis human kidney tissue labeling P Glycoprotein with unpurified ab170904 at 1/250 dilution.

Blocking and diluting buffer: 5% NFDM/TBST

Exposure time: Left image -10 seconds; Right image - 1 minute.

We suggest not to boil the sample after lysis.

For 1% SDS Hot Lysates preparation protocol, please refer to the protocol book in the protocol section and/or here (downloadable copy).

Blocking and diluting buffer: 5% NFDM/TBST

Exposure time: Left image - 3 seconds; Right image - 10 seconds.

We suggest not to boil the sample after lysis.

Blocking/Dilution buffer: 5% NFDM/TBST.

Blocking/Dilution buffer: 5% NFDM/TBST.

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

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

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human brain tissue labeling P Glycoprotein with unpurified ab170904 at 1/20. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody.

Counterstained with hematoxylin.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human brain tissue labeling P Glycoprotein with purified ab170904 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody.

Counterstained with hematoxylin.

P Glycoprotein antibody ab170904  was used with 3D Cell Culture Clearing Kit ab243299 to penetrate, stain and clear a 3D hepatocyte cell culture.

Blue: DAPI, Red:

Learn more about 3D cell culture and tissue clearing kits, reagents, and protocols designed to make it easier to stain 3D cell cultures and thick tissue sections and get more data from each valuable tissue section.