Recombinant Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker (ab154860)

Lanes 1-4: Merged signal (red and green). Green - ab154860 observed at 71 kDa. Red - loading control ab8245 observed at 36 kDa. 

 ab154860 Anti-Alas1 antibody [EPR10247]  was shown to specifically react with Alas1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab266473 (knockout cell lysate ab257348) was used.  Wild-type and Alas1 knockout samples were subjected to SDS-PAGE.  ab154860 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution 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 HepG2 cells labelling Alas1 with purified ab154860 at a dilution of 1/100. 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 (1/100) 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).

Immunocytochemistry/Immunofluorescence analysis of HepG2 cells labeling Alas1 with unpurified ab154860 at a dilution of 1/250.