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Palbociclib

Manufactured by Cayman Chemical
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Palbociclib is a selective cyclin-dependent kinase 4 and 6 (CDK4/6) inhibitor. It is used as a laboratory research tool to study cell cycle regulation and proliferation.

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Lab products found in correlation

Pyrazofurin, by Merck Group (2 mentions) Deoxycytidine, by Merck Group (2 mentions) Deoxythymidine, by Merck Group (2 mentions) Uridine, by Merck Group (2 mentions) Lometrexol, by Cayman Chemical (2 mentions) 6 azauridine, by Merck Group (2 mentions) Brequinar, by Merck Group (2 mentions) Tunicamycin, by Merck Group (2 mentions) Bay 2402234, by Bayer (2 mentions) Doxorubicin, by Merck Group (2 mentions) Rapamycin, by Merck Group (2 mentions) Chloroquine, by Merck Group (2 mentions) Puromycin, by Thermo Fisher Scientific (2 mentions) Doxycyclin, by Merck Group (2 mentions) Flavopiridol, by Cayman Chemical (2 mentions) Mk 1775, by Cayman Chemical (2 mentions) Zeocin, by Thermo Fisher Scientific (2 mentions) Bafilomycin a1, by Merck Group (2 mentions) 3 methyladenine, by Merck Group (2 mentions) Cisplatin, by Merck Group (2 mentions)

11 protocols using palbociclib

1

Nafamostat Mesylate Cytotoxicity Assay

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Cell cytotoxicity was determined using the 3-(4,5-dimethylthiazol-2-yl)−2,5-diphenyltetrazolium bromide (MTT) assay. For the effect of nafamostat mesylate on MCF7-TamR, MCF7-FulR, MCF7 cell lines were seeded in 96-well plates, treated with various concentrations of nafamostat mesylate (30, 60, and 90 μM) for 72–96 h. The medium was removed, and 30 μL of MTT (Sigma-Aldrich, St. Louis, MO, USA) at 0.5 mg/mL was added to the wells and then incubated at 37 °C for 3–4 h. The blue formazan crystals that formed were dissolved in DMSO, and the absorption was measured at 595 nm using an ELISA plate reader. For the combinational effect of nafamostat mesylate with CDK4/6 inhibitors palbociclib [Cayman Chemical (East Ellsworth Rd., MI, USA)], MCF7-TamR and MCF7-FulR cells were seeded in 96-well plates overnight followed by treatment with nafamostat mesylate (20 μM) and palbociclib (10 μM) for 72–96 h. The following process was described similarly as above.
Lin Y.T., Lin J., Liu Y.E., Hsu K.W., Hsieh C.C., Chen D.R, & Wu H.T. (2021). Nafamostat mesylate overcomes endocrine resistance of breast cancer through epigenetic regulation of CDK4 and CDK6 expression. Translational Oncology, 15(1), 101302.
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2

Palbociclib Antiproliferative Assay in Breast Cancer

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CTC‐ITB‐01 or MCF‐7 cells were seeded in a density of 2 × 103 cells per well in technical triplicates into a flat bottom 96‐well plate and placed in the incubator at 37°C with 5% CO2 overnight to adhere. The next day, Palbociclib (PD‐0332991, Cayman Chemical), dissolved in DMSO, was serially diluted to the desired concentration in standard cell culture medium and added to the cells. Afterward, the plate was immediately transferred into the IncuCyte Zoom live cell imaging system (Essen Bioscience) and relative cell confluence was measured every 2 h. The processing definition and confluence mask of the IncuCyte system were created using the provided IncuCyte Zoom 2016B software (Essen Bioscience). Accuracy of fit of the confluence mask was verified on representative images at different states of growth during the experiment.
Statistical testing was performed with Prism 7.0a (GraphPad Software Inc.) using a one‐way ANOVA followed by Dunnett's multiple comparisons test. A P < 0.05 was considered as statistically significant. IC50 calculation was also achieved with Prism 7.0a using the implemented non‐linear three‐parameter logistic curve regression. Concentrations were transformed to common logarithm.
Koch C., Kuske A., Joosse S.A., Yigit G., Sflomos G., Thaler S., Smit D.J., Werner S., Borgmann K., Gärtner S., Mossahebi Mohammadi P., Battista L., Cayrefourcq L., Altmüller J., Salinas‐Riester G., Raithatha K., Zibat A., Goy Y., Ott L., Bartkowiak K., Tan T.Z., Zhou Q., Speicher M.R., Müller V., Gorges T.M., Jücker M., Thiery J., Brisken C., Riethdorf S., Alix‐Panabières C, & Pantel K. (2020). Characterization of circulating breast cancer cells with tumorigenic and metastatic capacity. EMBO Molecular Medicine, 12(9), e11908.
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3

Synthesis of (R)-2-Hydroxyglutarate Ester

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TFMB ester of (R)-2-hydroxyglutarate was generated as previously described by R. Looper at University of Utah (Losman et al., 2013 (link)). BAY 2402234 was provided by Bayer Pharmaceuticals. Where indicated, cell culture media also contained the following additives: brequinar (Sigma-Aldrich), 6-azauridine (Sigma-Aldrich), pyrazofurin (Sigma-Aldrich), uridine (Sigma-Aldrich), lometrexol (Cayman), AGI-5198 (XcessBio), deoxythymidine (Sigma-Aldrich), deoxycytidine (Sigma-Aldrich), palbociclib (Cayman), tunicamycin (Sigma-Aldrich).
Shi D.D., Savani M.R., Levitt M.M., Wang A.C., Endress J.E., Bird C.E., Buehler J., Stopka S.A., Regan M.S., Lin Y.F., Puliyappadamba V.T., Gao W., Khanal J., Evans L., Lee J.H., Guo L., Xiao Y., Xu M., Huang B., Jennings R.B., Bonal D.M., Martin-Sandoval M.S., Dang T., Gattie L.C., Cameron A.B., Lee S., Asara J.M., Kornblum H.I., Mak T.W., Looper R.E., Nguyen Q.D., Signoretti S., Gradl S., Sutter A., Jeffers M., Janzer A., Lehrman M.A., Zacharias L.G., Mathews T.P., Losman J.A., Richardson T.E., Cahill D.P., DeBerardinis R.J., Ligon K.L., Xu L., Ly P., Agar N.Y., Abdullah K.G., Harris I.S., Kaelin WG J.r, & McBrayer S.K. (2022). De novo pyrimidine synthesis is a targetable vulnerability in IDH mutant glioma. Cancer cell, 40(9), 939-956.e16.
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4

Quantifying Breast Cancer Cell Viability

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For viability assays, we seeded cells as described above and incubated cells with various compounds or nutrient conditions for three days in LG/LF medium. We utilized compounds oligomycin A (#11342, Cayman Chemicals), fulvestrant (#S1191, Selleckchem, Houston, TX, USA), palbociclib (PD 0332991, #16273, Cayman Chemical), and syrosingopine (#SML1908, Millipore Sigma, Burlington, MA, USA). For different nutrient conditions, we used 2-deoxyglucose (2DG) (#14325, Cayman Chemical), sodium lactate (#L7022, Sigma Aldrich), and galactose (#G-0625, Sigma Aldrich). To quantify changes in relative numbers of viable breast cancer and stromal cells in co-cultures, we performed dual-color bioluminescence imaging of CBG (cancer cells) or CBRed (stromal cells) using an IVIS Lumina Series III (Perkin Elmer, Waltham, MA, USA) as previously described (18 ). Both CBG and CBR are ATP-dependent luciferase enzymes, so bioluminescence measures relative numbers of metabolically active cells. We exported photon flux data from regions of interest defined in Living Image 4.3.1 (Perkin Elmer) and quantified changes in bioluminescence signal with custom MATLAB code (5 ). We expressed data as percent change in bioluminescence relative to vehicle control, calculated as (treated bioluminescence – vehicle bioluminescence)/vehicle bioluminescence * 100.
Buschhaus J.M., Rajendran S., Chen S., Wharram B.L., Bevoor A.S., Cutter A.C., Humphries B.A., Robison T.H., Farfel A.P, & Luker G.D. (2023). Bone marrow mesenchymal stem cells induce metabolic plasticity in estrogen receptor-positive breast cancer. Molecular cancer research : MCR, 21(5), 458-471.
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5

Multi-drug Pharmacological Inhibition

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Doxycyclin (D9891), Doxorubicin (D1515), MG-132 (M7449), 3-Methyladenine (M9281), Chloroquine (C6628), Bafilomycin A1 (B1793), Cisplatin (1134357) and rapamycin (R8781) were purchased from Sigma-Aldrich (St Louis, MO, USA). Puromycin and Zeocin were purchased from Thermo Fisher Scientific (Grand Island, NY, USA). Cell cycle blockers, including MK-1775 (21266), Flavopiridol (10009197), Palbociclib (16273), were purchased from Cayman Chemical Company.
Zhang J., Tan P., Guo L., Gong J., Ma J., Li J., Lee M., Fang S., Jing J., Johnson G., Sun D., Cao W.M., Dashwood R., Han L., Zhou Y., Dong W.G, & Huang Y. (2018). p53-dependent autophagic degradation of TET2 modulates cancer therapeutic resistance. Oncogene, 38(11), 1905-1919.
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6

Adenoviral Vectors for IFNα2b Delivery

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Research-grade human serotype 5 adenoviral vectors lacking the E1 and E3 regions were produced by Viraquest Inc. Ad-IFNα2b is a dual expression vector with cytomegalovirus (CMV)-hIFNα2b in the E1 region and RSV-GFP in the E3 region. The Ad-GFP control vector has an empty E1 region and RSV-GFP in the E3 region (Figure 2A). The following other reagents were used in this study: rhIFNα2b (Stem Cell Technologies, 78,077), palbociclib (Cayman, 16,273), rhTRAIL (R&D 375-TL).
Green J.L., Osterhout R.E., Klova A.L., Merkwirth C., McDonnell S.R., Zavareh R.B., Fuchs B.C., Kamal A, & Jakobsen J.S. (2021). Molecular characterization of type I IFN-induced cytotoxicity in bladder cancer cells reveals biomarkers of resistance. Molecular Therapy Oncolytics, 23, 547-559.
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7

Multi-drug Pharmacological Inhibition

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Doxycyclin (D9891), Doxorubicin (D1515), MG-132 (M7449), 3-Methyladenine (M9281), Chloroquine (C6628), Bafilomycin A1 (B1793), Cisplatin (1134357) and rapamycin (R8781) were purchased from Sigma-Aldrich (St Louis, MO, USA). Puromycin and Zeocin were purchased from Thermo Fisher Scientific (Grand Island, NY, USA). Cell cycle blockers, including MK-1775 (21266), Flavopiridol (10009197), Palbociclib (16273), were purchased from Cayman Chemical Company.
Zhang J., Tan P., Guo L., Gong J., Ma J., Li J., Lee M., Fang S., Jing J., Johnson G., Sun D., Cao W.M., Dashwood R., Han L., Zhou Y., Dong W.G, & Huang Y. (2018). p53-dependent autophagic degradation of TET2 modulates cancer therapeutic resistance. Oncogene, 38(11), 1905-1919.
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8

Modulation of Hedgehog Signaling

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The following small molecules were used and dissolved in DMSO, ethanol, or water according to product data sheets as 1,000× stocks: cyclopamine (Cayman Chemical), SAG (Cayman chemical), Pr-8-Bromo cAMPs (sodium salt; Cayman Chemical), purmorphamine (Stemgent), GSA-10 (Tocris), palbociclib (Cayman Chemical), Torin 1, Ciliobrevin A (Tocris), and forskolin (Tocris). Small molecules were added to the cells 2 h before Hh induction and remained in the Shh CM throughout the experiment unless otherwise indicated.
Akhshi T, & Trimble W.S. (2020). A non-canonical Hedgehog pathway initiates ciliogenesis and autophagy. The Journal of Cell Biology, 220(1), e202004179.
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9

Physicochemical Properties of Enzastaurin and Palbociclib

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The compounds, namely, Enzastaurin and Palbociclib were purchased commercially from Cayman chemicals. The physicochemical properties of the naturally derived compounds Enzastaurin and Palbociclib were calculated using the Swiss ADME server [18 (link)].
Polisety A., Misra G., Rajawat J., Katiyar A., Singh H, & Bhatt A.N. (2022). Therapeutic natural compounds Enzastaurin and Palbociclib inhibit MASTL kinase activity preventing breast cancer cell proliferation. Medical Oncology (Northwood, London, England), 39(8), 100.
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10

Synthesis of (R)-2-Hydroxyglutarate Ester

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TFMB ester of (R)-2-hydroxyglutarate was generated as previously described by R. Looper at University of Utah (Losman et al., 2013 (link)). BAY 2402234 was provided by Bayer Pharmaceuticals. Where indicated, cell culture media also contained the following additives: brequinar (Sigma-Aldrich), 6-azauridine (Sigma-Aldrich), pyrazofurin (Sigma-Aldrich), uridine (Sigma-Aldrich), lometrexol (Cayman), AGI-5198 (XcessBio), deoxythymidine (Sigma-Aldrich), deoxycytidine (Sigma-Aldrich), palbociclib (Cayman), tunicamycin (Sigma-Aldrich).
Shi D.D., Savani M.R., Levitt M.M., Wang A.C., Endress J.E., Bird C.E., Buehler J., Stopka S.A., Regan M.S., Lin Y.F., Puliyappadamba V.T., Gao W., Khanal J., Evans L., Lee J.H., Guo L., Xiao Y., Xu M., Huang B., Jennings R.B., Bonal D.M., Martin-Sandoval M.S., Dang T., Gattie L.C., Cameron A.B., Lee S., Asara J.M., Kornblum H.I., Mak T.W., Looper R.E., Nguyen Q.D., Signoretti S., Gradl S., Sutter A., Jeffers M., Janzer A., Lehrman M.A., Zacharias L.G., Mathews T.P., Losman J.A., Richardson T.E., Cahill D.P., DeBerardinis R.J., Ligon K.L., Xu L., Ly P., Agar N.Y., Abdullah K.G., Harris I.S., Kaelin WG J.r, & McBrayer S.K. (2022). De novo pyrimidine synthesis is a targetable vulnerability in IDH mutant glioma. Cancer cell, 40(9), 939-956.e16.
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