Anti-VRK1 antibody [5D1] - N-terminal

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-VRK1 antibody [5D1] - N-terminal (AB171933)

Immunofluorescent analysis of formaldehyde-fixed interphase U-2 OS cells labeling VRK1 with ab171933 at 1/100 dilution (center). DNA was stained with DAPI (left) and two images were merged (right; merge).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-VRK1 antibody [5D1] - N-terminal (AB171933)

Immunofluorescent analysis of methanol-fixed metaphase U-2 OS cells labeling VRK1 with ab171933 at 1/100 dilution (center). DNA was stained with DAPI (left) and two images were merged (right; merge).

Metaphase cells. At metaphase, VRK1 dots were solely detected in nuclei.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-VRK1 antibody [5D1] - N-terminal (AB171933)

Immunofluorescent analysis of methanol-fixed interphase U-2 OS cells labeling VRK1 with ab171933 at 1/100 dilution (center). DNA was stained with DAPI (left) and two images were merged (right; merge).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-VRK1 antibody [5D1] - N-terminal (AB171933)

Immunofluorescent analysis of methanol-fixed interphase HeLa cells labeling VRK1 with ab171933 at 1/100 dilution (center). DNA was stained with DAPI (left) and two images were merged (right; merge).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-VRK1 antibody [5D1] - N-terminal (AB171933)

Immunofluorescent analysis of paraformaldehyde-fixed interphase HeLa cells labeling VRK1 with ab171933 at 1/100 dilution (center). DNA was stained with DAPI (left) and two images were merged (right; merge).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-VRK1 antibody [5D1] - N-terminal (AB171933)

Immunofluorescent analysis of methanol-fixed metaphase HeLa cells labeling VRK1 with ab171933 at 1/100 dilution (center). DNA was stained with DAPI (left) and two images were merged (right; merge).

Metaphase cells. At metaphase, VRK1 dots were solely detected in nuclei.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-VRK1 antibody [5D1] - N-terminal (AB171933)

Immunofluorescent analysis of paraformaldehyde-fixed metaphase HeLa cells labeling VRK1 with ab171933 at 1/100 dilution (center). DNA was stained with DAPI (left) and two images were merged (right; merge).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-VRK1 antibody [5D1] - N-terminal (AB171933)

Immunofluorescent analysis of formaldehyde-fixed metaphase U-2 OS cells labeling VRK1 with ab171933 at 1/100 dilution (center). DNA was stained with DAPI (left) and two images were merged (right; merge).

Metaphase cells. At metaphase, VRK1 dots were solely detected in nuclei.

• WB

Supplier Data

Western blot - Anti-VRK1 antibody [5D1] - N-terminal (AB171933)

All lanes:

Western blot - Anti-VRK1 antibody [5D1] - N-terminal (ab171933) at 1/500 dilution

Lane 1:

Extracts from TIG1 (5x10e4) cells: Luciferase RNAi (control)

Lane 2:

Extracts from TIG1 (5x10e4) cells: VRK1-1 RNAi

Lane 3:

Extracts from TIG1 (5x10e4) cells: VRK1-2 RNAi

Predicted band size: 45 kDa

true

• WB

Lab

Western blot - Anti-VRK1 antibody [5D1] - N-terminal (AB171933)

Lanes 1-3 : Merged signal (red and green). Green - ab171933 observed at 50 kDa. Red - loading control ab181602 observed at 36 kDa.

ab171933 Anti-VRK1 antibody [5D1] - N-terminal was shown to specifically react with VRK1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266257 (knockout cell lysate ab258283) was used. Wild-type and VRK1 knockout samples were subjected to SDS-PAGE. ab171933 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye^® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-VRK1 antibody [5D1] - N-terminal (ab171933) at 1/500 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

VRK1 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human VRK1 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-vrk1-knockout-hek-293t-cell-line-ab266257'>ab266257</a>)

Lane 3:

HeLa cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/10000 dilution

Predicted band size: 45 kDa

Observed band size: 50 kDa

false

• WB

Supplier Data

Western blot - Anti-VRK1 antibody [5D1] - N-terminal (AB171933)

Lanes 1 and 4:

Western blot - Anti-VRK1 antibody [5D1] - N-terminal (ab171933) at 1/100 dilution

Lanes 2 and 5:

Western blot - Anti-VRK1 antibody [5D1] - N-terminal (ab171933) at 1/500 dilution

Lanes 3 and 6:

Western blot - Anti-VRK1 antibody [5D1] - N-terminal (ab171933) at 1/1000 dilution

Lanes 1 - 3:

HeLa cell extract (5x10e4 cells)

Lanes 4 - 6:

U2OS cell extract (5x10e4 cells)

Secondary

All lanes:

Alexa488 goat anti-mouse IgG

Predicted band size: 45 kDa

true

• WB

Supplier Data

Western blot - Anti-VRK1 antibody [5D1] - N-terminal (AB171933)

All lanes:

Western blot - Anti-VRK1 antibody [5D1] - N-terminal (ab171933) at 1/500 dilution

Lane 1:

Extracts from HeLa (5x10e4) cells: Luciferase RNAi (control)

Lane 2:

Extracts from HeLa (5x10e4) cells: VRK1-1 RNAi

Lane 3:

Extracts from HeLa (5x10e4) cells: VRK1-2 RNAi

Predicted band size: 45 kDa

true

• WB

CiteAb

Western blot - Anti-VRK1 antibody [5D1] - N-terminal (AB171933)

VRK1 western blot using anti-VRK1 antibody [5D1] - N-terminal ab171933. Publication image and figure legend from Cartwright, T. N., Harris, R. J., et al., 2022, Sci Rep, PubMed 35778595.

ab171933 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab171933 please see the product overview.

RNAi studies of VRK kinase function in mitosis. (A) Kinome-wide RNAi screen for kinases involved in H3T3 phosphorylation in HeLa cells. The standard score reflects the intensity of H3T3ph. Note that negative values indicate a decline in H3T3ph intensity. The ranking of Haspin (red), VRK1, VRK2, VRK3 (blue) and PASK (black) are shown. Some results of this screen were previously published65. (B) RNAi depletion of VRK1 from HeLa cells does not cause a significant loss of H3T3ph as detected by immunoblot. The original blots are presented in Supplementary Fig. 7. (C) RNAi depletion of VRK1 does not cause a significant loss of H3T3ph or H3S10ph in individual mitotic HeLa cells, as detected by immunofluorescence microscopy. Scale bar, 10 µm. (D) Quantification of the proportion of HeLa cells in (C) with H3S10ph (i.e. mitotic cells) that also stain for H3T3ph.

false