Synthetic phospho-peptide corresponding to residues surrounding Tyr15 of human CDK1.
Species Reactivity
Human, Mouse
Validated Applications
WB, IHC-P
Molecular Weight
Predicted band size: 34 kDa
Positive Control
HeLa cell lysate, HeLa treated with 4mM hydroxyurea for 20 hours cell lysate, NIH/3T3 cell lysate, NIH/3T3 treated with 4mM hydroxyurea for 20 hours cell lysate, human breast cancer tissue.
Conjugation
unconjugated
Clone Number
PSH09-45
Reactivity Data
Tested Verified (internally validated)
Published Reported in literature (not internally validated)
Predicted Predicted reactive (based on sequence homology)
Not recommended Not recommended (failed internal validation)
WB
IHC-P
Human
Mouse
Rat
Product Features
Form
Liquid
Concentration
Lot Concentration Lookup
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Cyclin-dependent kinases (CDKs) are a predominant group of serine/threonine protein kinases involved in the regulation of the cell cycle and its progression, ensuring the integrity and functionality of cellular machinery. These regulatory enzymes play a crucial role in the regulation of eukaryotic cell cycle and transcription, as well as DNA repair, metabolism, and epigenetic regulation, in response to several extracellular and intracellular signals. They are present in all known eukaryotes, and their regulatory function in the cell cycle has been evolutionarily conserved. The catalytic activities of CDKs are regulated by interactions with CDK inhibitors (CKIs) and regulatory subunits known as cyclins. Cyclins have no enzymatic activity themselves, but they become active once they bind to CDKs. Without cyclin, CDK is less active than in the cyclin-CDK heterodimer complex. CDKs phosphorylate proteins on serine (S) or threonine (T) residues. The specificity of CDKs for their substrates is defined by the S/T-P-X-K/R sequence, where S/T is the phosphorylation site, P is proline, X is any amino acid, and the sequence ends with lysine (K) or arginine (R). This motif ensures CDKs accurately target and modify proteins, crucial for regulating cell cycle and other functions. Deregulation of the CDK activity is linked to various pathologies, including cancer, neurodegenerative diseases, and stroke.
Background References
1. Palacios-Blanco I et al. Cyclins and CDKs in the regulation of meiosis-specific events. Front Cell Dev Biol. 2022 Nov
2. Kohlmeyer JL et al. CDKs in Sarcoma: Mediators of Disease and Emerging Therapeutic Targets. Int J Mol Sci. 2020 Apr
Subcellular Location
Nucleus, Cytoplasm, Mitochondrion, cytoskeleton, microtubule organizing center, centrosome, spindle.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA723107) at 1/5,000 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.
☑ Cell treatment (CT)
Western blot analysis of Phospho-CDK (Y15) on different lysates with Rabbit anti-Phospho-CDK (Y15) antibody (HA723107) at 1/2,000 dilution.
Lane 1: NIH/3T3 cell lysate Lane 2: NIH/3T3 treated with 4mM hydroxyurea for 20 hours cell lysate Lane 3: NIH/3T3 treated with 4mM hydroxyurea for 20 hours cell lysate, then the membrane treated with λpp for 1 hour
Lysates/proteins at 20 µg/Lane.
Predicted band size: 34 kDa Observed band size: 34 kDa
Exposure time: 3 minutes; 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 (HA723107) 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.
☑ Cell treatment (CT)
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-Phospho-CDK (Y15) antibody (HA723107) 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 (HA723107) 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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