Recombinant Anti-CSN7b antibody [EPR6465] (ab124718)

Lanes 1-4: Merged signal (red and green). Green - ab124718 observed at 32 kDa. Red - loading control ab8245 observed at 36 kDa. 

 ab124718 Anti-CSN7b antibody [EPR6465]  was shown to specifically react with CSN7b in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266646 (knockout cell lysate ab257895) was used.  Wild-type and CSN7b knockout samples were subjected to SDS-PAGE.  ab124718 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.

Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: CSN7b knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: Jurkat cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab124718 observed at 32 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab124718 was shown to specifically react with CSN7b when CSN7b knockout samples were used. Wild-type and CSN7b knockout samples were subjected to SDS-PAGE. ab124718 and ab8245 (loading control to GAPDH) were both diluted 1/10000 and incubated overnight at 4°C. 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 1/10000 dilution for 1 hour at room temperature before imaging.

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling CSN7b with unpurified ab124718 at 1/800 dilution (1ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti-rabbit IgG (Alexa Fluor^® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.