MA1-828

antibody from Invitrogen Antibodies

Targeting: VRK1

Provider product page for MA1-828

Western blot

Immunocytochemistry

Immunohistochemistry

Antibody data

Antibody Data
Antigen structure
References [0]
Comments [0]
Validations
Western blot [2]
Immunocytochemistry [5]
Immunohistochemistry [3]

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Product number: MA1-828 - Provider product page
Provider: Invitrogen Antibodies
Product name: VRK1 Monoclonal Antibody (1F6)
Antibody type: Monoclonal
Antigen: Recombinant full-length protein
Description: MA1-828 detects VRK1 from human and mouse samples.
Antibody clone number: 1F6
Concentration: 1 mg/mL

VRK1 protein structure - MA1-828 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of HeLa (Lane 1), HEK 293 (Lane 2), MCF7 (Lane 3), and Jurkat (Lane 4). The blot was probed with Mouse Anti-VRK1 Monoclonal Antibody (Product # MA1-828, 1:500 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). A 48 kDa band corresponding to VRK1 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of VRK1 was performed by loading 25 µg of the indicated whole cell lysates and 5 µL of the Lane Marker Reducing Sample Buffer (Product # 39000) per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with a VRK1 monoclonal antibody (Product # MA1-828) at a dilution of 1:750 overnight at 4C on a rocking platform, washed in TBS-0.1%Tween 20, and probed with a goat anti-mouse IgG-HRP secondary antibody (Product # 32430) at a dilution of 1:10,000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34075).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of VRK1 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with VRK1 (1F6) Mouse Monoclonal Antibody (Product # MA1-828) at 1:100 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of VRK1 (green) in HeLa cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (left panel) or with (right panel) a VRK1 monoclonal antibody (Product # MA1-828) at a dilution of 1:200 overnight at 4°C, washed with PBS, and incubated with a DyLight-conjugated secondary antibody for 45 minutes at room temperature in the dark. F-actin (red) was stained with a fluorescent phalloidin and nuclei (blue) were stained with DAPI. Images were taken at a 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of VRK1 (green) in U251 cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (left panel) or with (right panel) a VRK1 monoclonal antibody (Product # MA1-828) at a dilution of 1:200 overnight at 4C, washed with PBS, and incubated with a DyLight-conjugated secondary antibody for 45 minutes at room temperature in the dark. F-actin (red) was stained with a fluorescent phalloidin and nuclei (blue) were stained with DAPI. Images were taken at a 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of VRK1 (green) in murine cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (left panel) or with (right panel) a VRK1 monoclonal antibody (Product # MA1-828) at a dilution of 1:200 overnight at 4°C, washed with PBS, and incubated with a DyLight-conjugated secondary antibody for 45 minutes at room temperature in the dark. F-actin (red) was stained with a fluorescent phalloidin and nuclei (blue) were stained with DAPI. Images were taken at a 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of VRK1 (green) in HeLa cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were then blocked with 5% normal goat serum (Product # 31873) for 15 minutes at room temperature. Cells were then probed with a VRK1 monoclonal antibody (Product # MA1-828) at a dilution of 1:100 for at least 1 hour at room temperature. Cells were then washed with PBS and incubated with DyLight 488 goat anti-mouse IgG secondary antibody (Product # 35502) at a dilution of 1:200 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on normal biopsies of deparaffinized human liver tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH 6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval, tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature and then probed without (left panel) or with (right panel) a VRK1 monoclonal antibody (Product # MA1-828) at a dilution of 1:200 overnight at 4C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and Streptavidin-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on normal biopsies of deparaffinized human testis tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH 6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval, tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature and then probed without (left panel) or with (right panel) a VRK1 monoclonal antibody (Product # MA1-828) at a dilution of 1:200 overnight at 4C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and Streptavidin-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on normal biopsies of deparaffinized human tonsil tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH 6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval, tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature and then probed without (left panel) or with (right panel) a VRK1 monoclonal antibody (Product # MA1-828) at a dilution of 1:200 overnight at 4C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and Streptavidin-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.