Anti-VRK1 antibody [5D1] - N-terminal (ab171933)

Mouse Monoclonal VRK1 antibody. N-terminal. Suitable for WB, ICC/IF and reacts with Human samples. Cited in 4 publications. Immunogen corresponding to Synthetic Peptide within Human VRK1 aa 1-50.

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Images

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

Publications

Key facts

Isotype: IgG1
Host species: Mouse
Storage buffer: pH: 6 - 8.5
Constituents: 50% Glycerol (glycerin, glycerine), 50% PBS
Form: Liquid
Clonality: Monoclonal

Immunogen

Synthetic Peptide within Human VRK1 aa 1-50. The exact immunogen used to generate this antibody is proprietary information. Database link Q99986

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	WB	ICC/IF
Human	Tested	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/200 - 1/1000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100	Notes -

Associated Products

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Target data

See full target information VRK1

Function

Serine/threonine kinase involved in the regulation of key cellular processes including the cell cycle, nuclear condensation, transcription regulation, and DNA damage response (PubMed:14645249, PubMed:18617507, PubMed:19103756, PubMed:33076429). Controls chromatin organization and remodeling by mediating phosphorylation of histone H3 on 'Thr-4' and histone H2AX (H2aXT4ph) (PubMed:31527692, PubMed:37179361). It also phosphorylates KAT5 in response to DNA damage, promoting KAT5 association with chromatin and histone acetyltransferase activity (PubMed:33076429). Is involved in the regulation of cell cycle progression of neural progenitors, and is required for proper cortical neuronal migration (By similarity). Is involved in neurite elongation and branching in motor neurons, and has an essential role in Cajal bodies assembly, acting through COIL phosphorylation and the control of coilin degradation (PubMed:21920476, PubMed:31090908, PubMed:31527692). Involved in Golgi disassembly during the cell cycle: following phosphorylation by PLK3 during mitosis, it is required to induce Golgi fragmentation (PubMed:19103756). Phosphorylates BANF1: disrupts its ability to bind DNA, reduces its binding to LEM domain-containing proteins and causes its relocalization from the nucleus to the cytoplasm (PubMed:16495336). Phosphorylates TP53BP1 and p53/TP53 on 'Thr-18', preventing the interaction between p53/TP53 and MDM2 (PubMed:10951572, PubMed:31527692). Phosphorylates ATF2 which activates its transcriptional activity (PubMed:15105425). Phosphorylates JUN (PubMed:31527692).

Alternative names

Serine/threonine-protein kinase VRK1, Vaccinia-related kinase 1, VRK1

Recommended products

Mouse Monoclonal VRK1 antibody. N-terminal. Suitable for WB, ICC/IF and reacts with Human samples. Cited in 4 publications. Immunogen corresponding to Synthetic Peptide within Human VRK1 aa 1-50.

Key facts

Isotype: IgG1
Host species: Mouse
Storage buffer: pH: 6 - 8.5
Constituents: 50% Glycerol (glycerin, glycerine), 50% PBS
Form: Liquid
Clonality: Monoclonal
Immunogen: Synthetic Peptide within Human VRK1 aa 1-50. The exact immunogen used to generate this antibody is proprietary information. Database link Q99986
Clone number: 5D1
Light chain type: kappa
Concentration
Purification notes: ab171933 was produced in serum-free medium and purified by propriety chromatography under mild conditions (90~98% pure).

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Product promise

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13 product images

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 Anti-GAPDH antibody [EPR16891] - 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 Human VRK1 knockout HEK-293T cell line ab266257 (knockout cell lysate Human VRK1 knockout HEK-293T cell lysate ab258283) was used. Wild-type and VRK1 knockout samples were subjected to SDS-PAGE. ab171933 and Anti-GAPDH antibody[EPR16891] - Loading Control (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 (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) and Goat anti-Mouse IgG H&L (IRDye^® 800CW) preadsorbed (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 (Human VRK1 knockout HEK-293T cell line ab266257)
Lane 3: HeLa cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/10000 dilution
Predicted band size: 45 kDa
Observed band size: 50 kDa
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).
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
Developed using the ECL technique.
Predicted band size: 45 kDa
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
Developed using the ECL technique.
Predicted band size: 45 kDa
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
Developed using the ECL technique.
Predicted band size: 45 kDa
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).
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).
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.
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).
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.
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).
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.
Image collected and cropped by CiteAb under a CC-BY license from the publication
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.