Phospho-CDK1 (T161) Recombinant Rabbit Monoclonal Antibody [PSH03-42]
cat.: HA721987
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IF-Cell, IHC-P, FC |
| Clonality: | Monoclonal |
| Clone number: | PSH03-42 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| 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: 34 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phosphopeptide corresponding to residues surrounding Thr161 of CDK1. |
| Positive control: | HeLa cell lysate, HeLa treated with 100nM Calyculin A for 30 minutes cell lysate, Jurkat cell lysate, Jurkat treated with 100nM Calyculin A for 30 minutes cell lysate, NIH/3T3 cell lysate, NIH/3T3 treated with 100nM Calyculin A for 30 minutes cell lysate, PC-12 cell lysate, PC-12 treated with UV for 1 hour cell lysate, human cervix cancer tissue, Jurkat cells treated with 100nM Calyculin A for 30 minutes, HeLa cells treated with 100nM Calyculin A for 30 minutes, NIH/3T3 cells treated with 100nM Calyculin A for 30 minutes. |
| Subcellular location: | Nucleus, Mitochondrion, Cytoplasm, cytoskeleton, microtubule organizing center, spindle. |
| Recommended Dilutions:
WB IF-Cell IHC-P FC |
1:2,000 1:2,500 1:200 1:100 |
| Uniprot #: | SwissProt: P06493 Human | P11440 Mouse | P39951 Rat |
| Alternative names: | Cdc 2 Cdc2 CDC28A CDK 1 CDK1 CDK1_HUMAN CDKN1 CELL CYCLE CONTROLLER CDC2 Cell division control protein 2 Cell division control protein 2 homolog Cell division cycle 2 G1 to S and G2 to M Cell division protein kinase 1 Cell Divsion Cycle 2 Protein Cyclin Dependent Kinase 1 Cyclin-dependent kinase 1 DKFZp686L20222 MGC111195 p34 Cdk1 p34 protein kinase P34CDC2 |
Images
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Fig1:
Western blot analysis of Phospho-CDK1 (T161) on different lysates with Rabbit anti-Phospho-CDK1 (T161) antibody (HA721987) at 1/2,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa treated with 100nM Calyculin A for 30 minutes cell lysate Lane 3: Jurkat cell lysate Lane 4: Jurkat treated with 100nM Calyculin A for 30 minutes cell lysate Lane 5: NIH/3T3 cell lysate Lane 6: NIH/3T3 treated with 100nM Calyculin A for 30 minutes cell lysate Lane 7: PC-12 cell lysate Lane 8: PC-12 treated with UV for 1 hour cell lysate Lysates/proteins at 30 µg/Lane. Predicted band size: 34 kDa Observed band size: 34 kDa Exposure time: 5 minutes; 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 (HA721987) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Immunohistochemical analysis of paraffin-embedded human cervix cancer tissue with Rabbit anti-Phospho-CDK1 (T161) antibody (HA721987) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (HA721987) 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. |
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Fig3:
Immunocytochemistry analysis of Jurkat cells treated with or without 100nM Calyculin A for 30 minutes labeling Phospho-CDK1 (T161) with Rabbit anti-Phospho-CDK1 (T161) antibody (HA721987) at 1/2,500 dilution. Cells were fixed in 100% precooled methanol for 5 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-Phospho-CDK1 (T161) antibody (HA721987) at 1/2,500 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 (M1305-2, 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. |
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Fig4:
Flow cytometric analysis of HeLa cells treated with or without 100nM Calyculin A for 30 minutes labeling Phospho-CDK1 (T161). Cells were fixed and permeabilized. Then stained with the primary antibody (HA721987, 1/100) (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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Fig5:
Flow cytometric analysis of NIH/3T3 cells treated with or without 100nM Calyculin A for 30 minutes labeling Phospho-CDK1 (T161). Cells were fixed and permeabilized. Then stained with the primary antibody (HA721987, 1/100) (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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