Anti-PBK/SPK antibody [EP2520Y] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBK/SPK antibody [EP2520Y] - BSA and Azide free (AB247421)

Immunohistochemical analysis of paraffin-embedded human tonsil using ab75987 at 1/100 dilution. Antigen retrieval was heat mediated with citrate buffer pH 6 before commencing with IHC staining protocol.

This data was developed using the same antibody clone in a different buffer formulation (ab75987).

• WB

Supplier Data

Western blot - Anti-PBK/SPK antibody [EP2520Y] - BSA and Azide free (AB247421)

This data was developed using the same antibody clone in a different buffer formulation (ab75987).

All lanes:

Western blot - Anti-PBK/SPK antibody [EP2520Y] - BSA and Azide free (ab247421) at 1/500000 dilution

All lanes:

HeLa cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 36 kDa

Observed band size: 40 kDa

false

• WB

Lab

Western blot - Anti-PBK/SPK antibody [EP2520Y] - BSA and Azide free (AB247421)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75987).

Lanes 1- 2 : Merged signal (red and green). Green - ab75987 observed at 40 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

ab75987 was shown to react with PBK/SPK in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266827 (knockout cell lysate ab257575) was used. Wild-type HEK-293T and PBK knockout HEK-293T cell lysates were subjected to SDS-PAGE. ab75987 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 100000 dilution and a 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-PBK/SPK antibody [EP2520Y] (<a href='/en-us/products/primary-antibodies/pbk-spk-antibody-ep2520y-ab75987'>ab75987</a>) at 1/100000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

PBK knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human PBK (SPK) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-pbk-spk-knockout-hek-293t-cell-line-ab266827'>ab266827</a>)

Predicted band size: 27 kDa,36 kDa

Observed band size: 40 kDa

false

• WB

Lab

Western blot - Anti-PBK/SPK antibody [EP2520Y] - BSA and Azide free (AB247421)

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : PBK knockout HAP1 cell lysate (20 μg)
Lane 3 : HeLa cell lysate (20 μg)
Lane 4 : Human tonsil tissue lysate (20 μg)
Lanes 1 - 4 : Merged signal (red and green). Green - ab75987 observed at 40 kDa. Red - loading control, ab7291, observed at 52 kDa.

ab75987 was shown to recognize PBK when PBK knockout samples were used, along with additional cross-reactive bands. Wild-type and PBK knockout samples were subjected to SDS-PAGE. ab75987 diluted to 1/100,000 and ab7291 (loading control to alpha Tubulin) diluted to 1/10,000 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 1/10,000 dilution for 1 hour at room temperature before imaging.

This data was developed using the same antibody clone in a different buffer formulation (ab75987).

All lanes:

Western blot - Anti-PBK/SPK antibody [EP2520Y] (<a href='/en-us/products/primary-antibodies/pbk-spk-antibody-ep2520y-ab75987'>ab75987</a>)

Predicted band size: 36 kDa

false