Anti-CSNK2A1 antibody [EP1963Y]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-CSNK2A1 antibody [EP1963Y] (AB76040)

Intracellular Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling CSNK2A1 with purified ab76040 at 1 : 20 dilution (10 μg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as a isotype control. Cell without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.

• WB

Lab

Western blot - Anti-CSNK2A1 antibody [EP1963Y] (AB76040)

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.

All lanes:

Western blot - Anti-CSNK2A1 antibody [EP1963Y] (ab76040) at 1/500 dilution

Lane 1:

Wild-type HAP1 cell lysate at 20 µg

Lane 2:

CSNK2A1 knockout HAP1 cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

Jurkat cell lysate at 20 µg

Predicted band size: 45 kDa

false

• WB

Lab

Western blot - 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.

All lanes:

Western blot - Anti-CSNK2A1 antibody [EP1963Y] (ab76040) at 1/1000 dilution

Lane 1:

Wild-type HCT116 cell lysate at 20 µg

Lane 2:

CSNK2A1 knockout HCT116 cell lysate at 20 µg

Lane 2:

Western blot - Human CSNK2A1 knockout HCT116 cell line (<a href='/en-us/products/cell-lines/human-csnk2a1-knockout-hct116-cell-line-ab266864'>ab266864</a>)

Predicted band size: 45 kDa

Observed band size: 45 kDa

false

• WB

Unknown

Western blot - Anti-CSNK2A1 antibody [EP1963Y] (AB76040)

All lanes:

Western blot - Anti-CSNK2A1 antibody [EP1963Y] (ab76040) at 1/1000 dilution

All lanes:

HeLa cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 45 kDa

Observed band size: 45 kDa

false

• WB

Lab

Western blot - Anti-CSNK2A1 antibody [EP1963Y] (AB76040)

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

All lanes:

Western blot - Anti-CSNK2A1 antibody [EP1963Y] (ab76040) at 1/1000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 µg

Lane 2:

Mouse brain lysate at 15 µg

Lane 3:

Rat brian lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 45 kDa

Observed band size: 45 kDa

false

• WB

Lab

Western blot - Anti-CSNK2A1 antibody [EP1963Y] (AB76040)

ab76040 was shown to react with CSNK2A1 in wild-type HAP1 cells in Western blot with loss of signal observed in a CSNK2A1 knockout cell line. Wild-type HAP1 and CSNK2A1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab76040 overnight at 4 °C at a 1/500 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Western blot - Anti-CSNK2A1 antibody [EP1963Y] (ab76040) at 1/500 dilution

Lane 1:

Wild-type HAP1 lysate at 30 µg

Lane 2:

CSNK2A1 knock-out HAP1 lysate at 30 µg

false