Recombinant Anti-CSNK2A1 antibody [EP1963Y] (ab76040)

Lanes 1-2: Merged signal (red and green). Green - ab76040 observed at 45 kDa. Red - loading control ab8245 observed at 37 kDa.

 ab76040 Anti-CSNK2A1 antibody [EP1963Y] was shown to specifically react with CSNK2A1 in wild-type HCT116 cells. Loss of signal was observed when knockout cell line ab266864 (knockout cell lysate ab257401) was used. Wild-type and CSNK2A1 knockout samples were subjected to SDS-PAGE. ab76040 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 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.

Lanes 1 - 4: Merged signal (red and green). Green – ab76040 observed at 45 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab76040 was shown to specifically react with CSNK2A1 when CCSNK2A1 knockout samples were used. Wild-type and CSNK2A1 knockout samples were subjected to SDS-PAGE. ab76040 and ab8245 (loading control to GAPDH) were diluted at 1/500 and 1/10 000 respectively 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.

Immunohistochemical analysis of CSNK2A1 in paraffin embedded human breast carcinoma tissue using ab76040 at a 1/50 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Overlay histogram showing Jurkat cells stained with ab76040 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab76040, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.