Roscovitine

Manufactured by LC Laboratories

Sourced in United States

Roscovitine is a cyclin-dependent kinase (CDK) inhibitor used as a research tool. It acts by selectively inhibiting certain CDK enzymes, which play a role in cell cycle regulation. Roscovitine is commonly used in life science research applications to study cell cycle processes.

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Lab products found in correlation

Gefitinib, by LC Laboratories (2 mentions) Tozasertib, by LC Laboratories (2 mentions) Hdm201, by Novartis (1 mentions) Sk mel 5, by American Type Culture Collection (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Mcf 7, by American Type Culture Collection (1 mentions) Dmem f12 medium, by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Palbociclib, by LC Laboratories (1 mentions) Cgm097, by Novartis (1 mentions) Buparlisib, by Novartis (1 mentions) Ribociclib, by Novartis (1 mentions) Ci 1040, by Selleck Chemicals (1 mentions) Danusertib, by Selleck Chemicals (1 mentions) Sb218078, by Bio-Techne (1 mentions) Ku55933, by Selleck Chemicals (1 mentions) Plx4720, by Selleck Chemicals (1 mentions) Pf573228, by Bio-Techne (1 mentions) Plx4032, by Selleck Chemicals (1 mentions) Pf 4708671, by Selleck Chemicals (1 mentions)

6 protocols using roscovitine

Pharmacological Inhibition of CDK4/6 in Melanoma

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A375, SK-Mel5, MCF7 cells were purchased from ATCC. YUMM10.1 and YUMM1.7 cells were provided by Marcus Bosenberg (Yale University). Cells were cultured in DMEM/F12 medium supplemented with 10% FBS and 1% penicillin/streptomycin (all Gibco). HDM201, CGM097, ribociclib, and buparlisib were provided by Novartis. GDC-0994 and ipatasertib were purchased from Medkoo. Palbociclib and roscovitine were purchased from LC laboratories. Nutlin-3a was synthesized as described previously (70 (link)). Working concentrations of CDK4/6 inhibitors ribociclib and Palbociclib used for in vitro experiments were chosen based on previously reported human pharmacokinetics data and kinase-specificity assays (71 (link)-74 (link)). Antibodies used in this study are indicated in table S2.

Vilgelm A.E., Saleh N., Shattuck-Brandt R., Riemenschneider K., Slesur L., Chen S.C., Johnson C.A., Yang J., Blevins A., Yan C., Johnson D.B., Al-Rohil R.N., Halilovic E., Kauffmann R.M., Kelley M., Ayers G.D, & Richmond A. (2019). MDM2 Antagonists Overcome Intrinsic Resistance to CDK4/6 Inhibition by Inducing p21. Science translational medicine, 11(505), eaav7171.

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Screening of Protein Kinase Inhibitors for TNBC

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The fifty-five protein kinase inhibitors (PKIs) were purchased from following sources: BML-275, FR 180204, IKK16, GW 843682X, NSC 109555, NU7441, PD407824, PF 573228, SB 218078, TCS PIM-1-1, TCS PIM-1-4a, and TPCA-1 from Tocris Biosciences (Bristol, UK); indirubin-3′-monoxime and Ro-31-8220 from Calbiochem (San Diego, CA, USA); A-769662, bosutinib, chelerythrine, CP690550, fasudil, gefitinib, imatinib, nilotinib, PKC412, roscovitine, SNS-314, and tozasertib from LC Laboratories (Woburn, MA, USA); AT7867, AT9283, AZD1152, AZD1480, BI 2536, BIX 02189, CHIR-99021, CI-1040, CYC116, danusertib, enzastaurin, GDC-0879, INCB018424, JNJ-7706621, KU-55933, LY2228820, MLN8237, PD-0325901, PF-4708671, PLX-4032, PLX-4720, SB216763, SNS-032, SP600125, VX-702, Y-27632, and ZM447439 from Selleck Chemicals (Houston, TX, USA); U0126 from Promega (Madison, WI, USA); TBCA from Millipore (Burlington, MA, USA).
All TNBC cells in this study were obtained from the American Type Culture Collection (Manassas, VA, USA). The cultured cells were monitor by trypan blue cell counting as described previously [58 (link)].

Yi Y.W., You K.S., Han S., Ha I.J., Park J.S., Lee S.G, & Seong Y.S. (2022). Inhibition of IκB Kinase Is a Potential Therapeutic Strategy to Circumvent Resistance to Epidermal Growth Factor Receptor Inhibition in Triple-Negative Breast Cancer Cells. Cancers, 14(21), 5215.

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Prednisolone and Roscovitine on Murine Bone

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Eleven-week-old wild-type female BALB/cAnNCrl mice (Charles River Laboratories, Wilmington, MA, USA) received a subcutaneous slow-release sham- or prednisolone (Pred)-pellet (12 mg/kg/day) (Innovative Research of America, Sarasota, FL, USA) at the neck, as previously described [58 (link)]. The mice were injected intraperitoneally (i.p.) with either a vehicle (5% dimethyl sulfoxide (DMSO), 10% kolliphor EL (Sigma-Aldrich, Taufkirchen, Germany), 85% 1× PBS), or roscovitine (150 mg/kg) (LC Laboratories, Woburn, MA, USA), three times a week for two weeks, as previously described [53 (link)]. The surgery was performed under general anesthesia (2 volume percent (vol%) isoflurane (Baxter, Unterschleißheim, Germany)). After two weeks, the mice were euthanized by an overdose of isoflurane, and the femora were collected for further analyses.

Krüger B.T., Steppe L., Vettorazzi S., Haffner-Luntzer M., Lee S., Dorn A.K., Ignatius A., Tuckermann J, & Ahmad M. (2022). Inhibition of Cdk5 Ameliorates Skeletal Bone Loss in Glucocorticoid-Treated Mice. Biomedicines, 10(2), 404.

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Intrathecal Administration of MEK and Cdk5 Inhibitors for Pain Management

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Male Sprague-Dawley rats (220∼250 g) were obtained from Shanghai laboratory animal center (Shanghai, China). All experimental procedures were approved by the Committee of Animal Use for Research and Education of Shanghai Jiao Tong University School of Medicine. The numbers of animals used in all experiments were minimized according to the guidelines established by the Ethical Issues of the International Association for the study of pain. Rats were housed in a climate-controlled environment on a12 h light/dark cycles and free access to food and water. CFA (100 µl, Sigma, St. Louis, MO, USA) or 5% formalin (100 µl) was injected into the plantar surface of the left hind paw of rats. For intrathecal drug delivery, a polyethylene-10 catheter was implanted into the intrathecal space of the spinal cord at the lumbar enlargement, and 5 µl of the MEK inhibitor U0126 (2 µg, Sigma) dissolved in 4% dimethyl sulfoxide (DMSO), was administered 30 min before treatment and once per day (for 7 days) after treatment, DMSO (4%) was injected as vehicle control. 5 µl of the Cdk5 inhibitor roscovitine (100 µg, R-1234, LC Laboratories, Woburn, MA, USA) dissolved in 10% DMSO, was administered 30 min before treatment and once per day (for 7 days) after treatment, DMSO (10%) was injected as vehicle control.

Zhang X., Zhang H., Shao H., Xue Q, & Yu B. (2014). ERK MAP Kinase Activation in Spinal Cord Regulates Phosphorylation of Cdk5 at Serine 159 and Contributes to Peripheral Inflammation Induced Pain/Hypersensitivity. PLoS ONE, 9(1), e87788.

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Kinase Assay Protocol with Diverse Components

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Kinases, substrates, and cofactors were obtained from Signal-Chem (Richmond, BC, Canada), except PKA, PKC, DNA-PK, PI3 kinases a, b, g, and d, neurogranin, kemptide, and DNA-PK peptide, which were obtained from Promega (Madison, WI, USA). Table S1 in the Online Supplementary Material provides a list of all the kinases, substrates, and cofactors used in this study. SB203580 was obtained from Promega. Gefitinib, roscovitine, dasatinib, tofacitinib, tozasertib, LY294002, AZD6482, AS252424, and AS605240 were purchased from LC Laboratories (Woburn, MA, USA). The ADP-Glo kinase assay was obtained from Promega. Low-volume, 384-well, white round-bottom untreated polystyrene microplates and halfarea 96-well white untreated microplates were obtained from Corning (Corning, NY, USA). Bovine serum albumin (BSA; blotqualified BSA, cat. no. 3841) was obtained from Promega.

Hennek J., Alves J., Yao E., Goueli S., & Zegzouti H. (2015). Bioluminescent kinase strips: A novel approach to targeted and flexible kinase inhibitor profiling. Analytical Biochemistry, 495, 9-20.

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Glutamate-induced Cytotoxicity Assay

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Glutamate, 3-(4,5-dimethyldiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), and poly-l-lysine were obtained from Sigma (St. Louis, MO). Roscovitine was purchased from LC Laboratories (Woburn, MA). ALDH1A1 (L-15), Cdk5 (C-8), Cdk5 (DC-17), p35 (C-19), actin (C-2), alpha-tubulin (TU-02), lamin A (H-102), and enolase (N-14) were purchased from Santa Cruz Biotech (Santa Cruz, CA) (Supplementary File 1). All antibodies were used at 1:1000 dilution. Validation for ALDH1A1 and Cdk5 antibody has been shown in Supplementary Fig. 1. HT22 cells were a gift from David Schubert. HEK293T and Phoenix cells were purchased from ATCC (Manassas, VA). HT22, HEK293T, and Phoenix cells were cultured in DMEM with 10% FBS supplemented with 2 mM glutamine and antibiotics.

Nikhil K., Viccaro K, & Shah K. (2018). Multifaceted Regulation of ALDH1A1 by Cdk5 in Alzheimer’s Disease Pathogenesis. Molecular neurobiology, 56(2), 1366-1390.

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