CDC2/CDK1 [p Tyr15] Antibody [Unconjugated]

Western blot shows lysates of L-929 mouse fibroblast cell line and Jurkat human acute T cell leukemia cell line untreated (-) or treated (+) with 0.2 µg/mL nocodazole or 12 µM aphidicolin or 1 mM hydroxurea for 18 hours. PVDF membrane was probed with 0.2 µg/mL of Rabbit Anti-Human/Mouse/Rat Phospho-CDC2 (Y15) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF888), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-CDC2 (Y15) at approximately 34 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Cooperative induction of DNA damage in vivo by WEE1 and CHK1 inhibitors. LoVo xenograft tumor-bearing mice were treated with 60 mpk MK-1775 BID for 2 days, 60 mpk MK-8776 BID for 2 days, or the combination of MK-1775 and MK-8776 each at 60 mpk BID for 2 days. Tumors were collected at 2, 24, and 48 hours following the final dose. A, LoVo tumor lysates were analyzed by Western blot for pCHK1S345. B, Tumor sections were fixed and analyzed by immunohistochemistry (IHC). Representative images for gamma H2AX at 2 hours and 48 hours post final dose are shown. C, Quantitative analysis of IHC for both phospho-CHK1S345 and gamma H2AX (n=3); one-way ANOVA analyses *P<0.05. **P<0.01, ***P<0.001. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/23148684), licensed under a CC-BY license. Not internally tested by R&D Systems.

DNA damage response incurred by MK-1775 and MK-8776 is dependent on CDK activity.A, Resistant (H460) or sensitive (LoVo) cells were treated with concentrations of MK-1775 and MK-8776 described for Figure 4, or 1 uM nocodazole for control. After 24 hours, cells were harvested and lysates analyzed by Western blot for caspase-dependent cleaved PARP (PARP*). B, A2058, HT-29, and LoVo cells were treated for 30 minutes with either DMSO or the indicated concentration of CDK inhibitor (SCH-727965). Following this pretreatment, further DMSO or concentrations of MK-1775 and MK-8776 used in Figures 3 and 4 (125 nM MK-1775 plus 150 nM MK-8776 in A2058; 125 nM MK-1775 plus 300 nM MK-8776 in HT-29, and 40 nM MK-1775 plus 75 nM MK-8776 in LoVo) were added to the cells for an additional 2 hours before cells were harvested and lysates analyzed by Western blot for phosphorylated CHK1S345, indicative of activated DNA damage response. C, LoVo cells were treated for 2 hours with 75 nM MK-1775 alone or in combination with 150 nM MK-8776, as indicated. Cells were harvested and lysates analyzed by Western blot for the proteins and phosphoproteins indicated. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/23148684), licensed under a CC-BY license. Not internally tested by R&D Systems.

DNA damage response incurred by MK-1775 and MK-8776 is dependent on CDK activity.A, Resistant (H460) or sensitive (LoVo) cells were treated with concentrations of MK-1775 and MK-8776 described for Figure 4, or 1 uM nocodazole for control. After 24 hours, cells were harvested and lysates analyzed by Western blot for caspase-dependent cleaved PARP (PARP*). B, A2058, HT-29, and LoVo cells were treated for 30 minutes with either DMSO or the indicated concentration of CDK inhibitor (SCH-727965). Following this pretreatment, further DMSO or concentrations of MK-1775 and MK-8776 used in Figures 3 and 4 (125 nM MK-1775 plus 150 nM MK-8776 in A2058; 125 nM MK-1775 plus 300 nM MK-8776 in HT-29, and 40 nM MK-1775 plus 75 nM MK-8776 in LoVo) were added to the cells for an additional 2 hours before cells were harvested and lysates analyzed by Western blot for phosphorylated CHK1S345, indicative of activated DNA damage response. C, LoVo cells were treated for 2 hours with 75 nM MK-1775 alone or in combination with 150 nM MK-8776, as indicated. Cells were harvested and lysates analyzed by Western blot for the proteins and phosphoproteins indicated. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/23148684), licensed under a CC-BY license. Not internally tested by R&D Systems.

Product Discontinued

CDC2/CDK1 [p Tyr15] Antibody [Unconjugated] Summary

• Immunogen: Phosphopeptide containing CDC2 Y15 site
• Modification: p Tyr15
• Specificity: Detects human, mouse, and rat CDC2 when phosphorylated at Y15.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: CDK1
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 5-15 ug/mL
  • Western Blot 0.2 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for CDC2/CDK1 [p Tyr15] Antibody [Unconjugated]

• CDC2
• CDC28A
• CDC2MGC111195
• CDK1
• cell cycle controller CDC2
• Cell division control protein 2 homolog
• Cell division protein kinase 1
• cyclin-dependent kinase 1
• DKFZp686L20222
• EC 2.7.11.22
• EC 2.7.11.23
• G1 to S and G2 to M
• p34 protein kinase
• P34CDC2

Background

CDC2 (Cell Division Cycle 2), also known as CDK1 (Cyclin Dependent Kinase 1), is a member of the CDK family of serine/threonine kinases. The CDKs are important regulators of cell cycle progression and their activities are largely controlled by association with Cyclins, and activating and inhibitory phosphorylations. Entry into mitosis is initiated by CDC2. Full activation of CDC2 requires phosphorylation at T161 and association with CyclinB. In contrast, phosphorylation of CDC2 at Y15 and T14 during the G2-phase of the cell cycle inhibits activity, and dephosphorylation of Y15 and T14 by CDC25 phosphatase during late G2 restores activity.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for CDC2/CDK1 Antibody (AF888)(5)

Publications using AF888

• J Liu, Y Wang, C Mu, M Li, K Li, S Li, W Wu, L Du, X Zhang, C Li, W Peng, J Shen, Y Liu, D Yang, K Zhang, Q Ning, X Fu, Y Zeng, Y Ni, Z Zhou, Y Liu, Y Hu, X Zheng, T Wen, Z Li, Y Liu Pancreatic tumor eradication via selective Pin1 inhibition in cancer-associated fibroblasts and T lymphocytes engagement Nature Communications, 2022-07-25;13(1):4308. 2022-07-25 [PMID: 35879297] (Western Blot, Mouse)
• X Wang, Z Chen, AK Mishra, A Silva, W Ren, Z Pan, JH Wang Chemotherapy-induced differential cell cycle arrest in B cell lymphomas affects their sensitivity to Wee1 inhibition Haematologica, 2017-12-07;0(0):. 2017-12-07 [PMID: 29217775] (Western Blot, Mouse)
• Guertin , Amy D, Martin , Melissa, Roberts , Brian, Hurd , Melissa, Qu , Xianlu, Miselis , Nathan R, Liu , Yaping, Li , Jing, Feldman , Igor, Benita , Yair, Bloecher , Andrew, Toniatti , Carlo, Shumway , Stuart D Unique functions of CHK1 and WEE1 underlie synergistic anti-tumor activity upon pharmacologic inhibition. Cancer Cell Int, 2012-11-13;12(1):45. 2012-11-13 [PMID: 23148684] (Neutralization, Mouse)
• Jingyu Zhang, Luis A. Espinoza, Robert J. Kinders, Scott M. Lawrence, Thomas D. Pfister, Ming Zhou et al. NANOG modulates stemness in human colorectal cancer Oncogene 2013-09-12 [PMID: 23085761]
• Tcherpakov M, Bronfman FC, Conticello SG, Vaskovsky A, Levy Z, Niinobe M, Yoshikawa K, Arenas E, Fainzilber M The p75 neurotrophin receptor interacts with multiple MAGE proteins. J. Biol. Chem., 2002-10-31;277(51):49101-4. 2002-10-31 [PMID: 12414813] (Western Blot, Rat)

Bioinformatics

• Gene Symbol: CDK1