MSK1 Recombinant Rabbit Monoclonal Antibody [PSH13-72]
cat.: HA723561
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat, Monkey
Applications: WB, IF-Cell, IHC-P, FC
Clonality: Monoclonal
Clone number: PSH13-72
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 90 kDa
Isotype: IgG
Immunogen: Recombinant protein within human MSK1 aa 351-802.
Positive control: HeLa cell lysate, SK-Br-3 cell lysate, 293T cell lysate, L-929 cell lysate, C6 cell lysate, Mouse liver tissue lysate, COS-1 cell lysate, SK-Br-3, L-929, C6, human brain tissue, human colon tissue, mouse brain tissue, mouse colon tissue, mouse liver tissue, rat brain tissue, rat colon tissue, rat liver tissue.
Subcellular location: Nucleus, Cytoplasm.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
1:20,000-1:50,000
1:100
1:200-1:1,000
1:1,000
Uniprot #: SwissProt: O75582 Human | Q8C050 Mouse
Entrez Gene: 314384 Rat
Alternative names: 90 kDa ribosomal protein S6 kinase 5 EC 2.7.11.1 KS6A5_HUMAN MGC1911 Mitogen and stress activated protein kinase 1 MSPK1 Nuclear Mitogen And Stress Activated Protein Kinase 1 Nuclear mitogen- and stress-activated protein kinase 1 Ribosomal protein S6 kinase 90kD polypeptide 5 Ribosomal protein S6 kinase 90kDa Ribosomal protein S6 kinase 90kDa polypeptide 5 Ribosomal Protein S6 Kinase Alpha 5 Ribosomal protein S6 kinase alpha-5 RLPK RPS6KA5 RSK Like Protein Kinase RSK-like protein kinase RSKL S6K alpha 5 S6K-alpha-5
Images
HA723561_1.jpg Fig1: Western blot analysis of MSK1 on different lysates with Rabbit anti-MSK1 antibody (HA723561) at 1/20,000 dilution.

Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: SK-Br-3 cell lysate (20 µg/Lane)
Lane 3: 293T cell lysate (20 µg/Lane)
Lane 4: L-929 cell lysate (20 µg/Lane)
Lane 5: C6 cell lysate (20 µg/Lane)
Lane 6: Mouse liver tissue lysate (low expression) (30 µg/Lane)
Lane 7: COS-1 cell lysate (20 µg/Lane)

Predicted band size: 90 kDa
Observed band size: 90 kDa

Exposure time: 10 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA723561) at 1/20,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA723561_2.jpg Fig2: Immunocytochemistry analysis of SK-Br-3 cells labeling MSK1 with Rabbit anti-MSK1 antibody (HA723561) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-MSK1 antibody (HA723561) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA723561_3.jpg Fig3: Immunocytochemistry analysis of L-929 cells labeling MSK1 with Rabbit anti-MSK1 antibody (HA723561) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-MSK1 antibody (HA723561) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA723561_4.jpg Fig4: Immunocytochemistry analysis of C6 cells labeling MSK1 with Rabbit anti-MSK1 antibody (HA723561) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-MSK1 antibody (HA723561) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA723561_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-MSK1 antibody (HA723561) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723561) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723561_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-MSK1 antibody (HA723561) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723561) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723561_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-MSK1 antibody (HA723561) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723561) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723561_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-MSK1 antibody (HA723561) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723561) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723561_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-MSK1 antibody (HA723561) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723561) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723561_10.jpg Fig10: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-MSK1 antibody (HA723561) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723561) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723561_11.jpg Fig11: Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-MSK1 antibody (HA723561) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723561) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723561_12.jpg Fig12: Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-MSK1 antibody (HA723561) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723561) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723561_13.jpg Fig13: Flow cytometric analysis of SK-Br-3 cells labeling MSK1.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA723561, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
HA723561_14.jpg Fig14: Flow cytometric analysis of L-929 cells labeling MSK1.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA723561, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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