Recombinant Anti-MVP antibody [EPR13227(B)] (ab175239)

Lanes 1-4: Merged signal (red and green). Green - ab175239 observed at 110 kDa. Red - loading control ab8245 observed at 37 kDa.

 ab175239 Anti-MVP antibody [EPR13227(B)] was shown to specifically react with MVP in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264817 (knockout cell lysate ab257544) was used. Wild-type and MVP knockout samples were subjected to SDS-PAGE. ab175239 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 2000 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling MVP with purified ab175239 at a dilution of 1/150. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

Control 1: primary antibody (1/150) and secondary antibody, ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000).

Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue labelling MVP with purified ab175239 at a dilution of 1/350. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

Intracellular Flow Cytometry analysis of A549 cells labelling MVP with purified ab175239 at a dilution of 1/180 (red). Cells were fixed with 4% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies. 

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human pancreas tissue labeling MVP with unpurified ab175239 at a dilution of 1/50.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue labeling MVP with unpurified ab175239 at a dilution of 1/50.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Immunocytochemistry/Immunofluorescence analysis of A549 cells labeling MVP with unpurified ab175239 at a dilution of 1/250 (red) and DAPI staining (blue).

Intracellular flow cytometrical analysis of permeabilized A549 cells labeling MVP with unpurified ab175239 antibody at a dilution of 1/100 (red) compared to a negative control (Rabbit IgG, green).