Anti-VRK1 (human) antibody, mouse monoclonal (5D1)
Product code71-600

Size

100 µgStorage-20℃

Concentration

1.0 mg/ml

Buffer

PBS- with 50% glycerol

PurityPurified IgG fraction with protein A from hybridoma cell culture medium.

ImmunogenSynthetic peptide corresponding to N-terminus of human VRK1,MPRVKAAQAGRQSSAKRHL-C

Isotype

Mouse IgG1κ

Reactivity

Human VRK1 protein. Not tested with other species.

Special notes

N/A

Application

1. Western blotting (1/200~1/1,000 dilution). Use ofhighly sensitive

chemiluminescence reagents such as Lumi-Light Plus(Roche) or

R LD (Wako, Tokyo) are recommended.
ImmunoStar○

2. Immunoprecipitation (assay dependent)
3. Immunofluorescence staining (1/100 dilution)
4. Immunohistochemistry (assay dependent)
5. ELISA (assay dependent)
Background

The VRK1 gene encodes serine/threonine kinase VRK1 (Vaccinia-Related Kinase 1;
396 aa, 45.5 kDa) which is involved in Golgi disassembly during the cell cycle
following phosphorylation by PLK3 during mitosis, and required to induce Golgi
fragmentation. It acts by mediating phosphorylation of a downstream target protein'Thr-18' of p53/TP53 and may thereby prevent the interaction between p53/TP53
and MDM2. It also phosphorylates casein and histone H3. Phosphorylation of theBANF1 gene product disrupts its ability to bind DNA, reduces its binding to LEM
domain-containing proteins and causes its relocalization from the nucleus to thecytoplasm.
Involvement in disease

Defects in VRK1 are the cause of pontocerebellar

hypoplasia type 1A (PCH1A); also called pontocerebellar hypoplasia with infantile
spinal muscular atrophy or pontocerebellar hypoplasia with anterior horn celldisease. PCH1A is characterized by an abnormally small cerebellum and brainstem,
central and peripheral motor dysfunction from birth, gliosis and anterior horn celldegeneration resembling infantile spinal muscular atrophy.

Data Link

SwissProt: Q99986 Human , Entrez Gene: 7443 HumanPlease note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC
PROCEDURES. NOT FOR MILITARY USE.
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Data Images: 71-600 Anti-VRK1 (human) antibody, mouse monoclonal (5D1)

Fig.1

Western blot detection of VRK1 in the crude extracts of human cells. Lanes 1, 2, 3; HeLa cellextract (5x104 cells) with antibody dilutions at 1/100, 1/500, 1/1000. Lanes 4, 5, 6; U2OS cell extract(5x104 cells) with the antibody dilutions at 1/100, 1/500, 1/1,000. As secondary antibody, Alexa488
R LD (Wako, Tokyo) was used as chemiluminescencegoat anti-mouse IgG was used. ImmunoStar○reagent and images were taken with BIO-RAD ChemiDocXRS.

Fig.2. Inhibition of VRK1 expression in human cells treated by RNAi. specific to VRK1.
Lane 1; Luciferase RNAi (control). Lane2; VRK1-1 RNAi. Lane 3; VRK1-2 RNAi.. Antibody at 1/500
dilution. Lumi-Light Plus (Roche) was used as chemiluminescence reagent. Extracts from 5x104 cells.
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Fig.3. Immunoflorescence staining of VRK1 in HeLa cells, PA fixed.
Top; Interphase cells were fixed with paraformaldehyde and stained with the anti-human VRK1
antibody (hvrk1) at 1/100 dilution (center), DNA was stained with DAPI (left) and two images weremerged (right; merge).
Bottom; Metaphase cells. At metaphase, VRK1 dots were solely detected in nuclei.

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Fig.4. Immunoflorescence staining of VRK1 in HeLa cells, methanol fixed.
Top; Interphase cells were fixed with methanol and stained with the anti-human VRK1 antibody(hvrk1) at 1/100 dilution (center), DNA was stained with DAPI (left) and two images were merged(right; merge).
Center and bottom; Metaphase cells. At metaphase, VRK1 dots were solely detected in nuclei.

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Fig. 5. Immunoflorescence staining of VRK1 in U-2 OS cells, formaldehyde fixed.
Top; Interphase cells were fixed with formaldehyde and stained with the anti-human VRK1 antibody(hvrk1) at 1/100 dilution (center), DNA was stained with DAPI (left) and two images were merged(right; merge).
Center and bottom; Metaphase cells. At metaphase, VRK1 dots were solely detected in nuclei.

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Fig. 6. Immunoflorescence staining of VRK1 in U-2 OS cells, methanol fixed.
Top; Interphase cells were fixed with methanol and stained with the anti-human VRK1 antibody(hvrk1) at 1/100 dilution (center), DNA was stained with DAPI (left) and two images were merged(right; merge).
Center and bottom; Metaphase cells. At metaphase, VRK1 dots were solely detected in nuclei.

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