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Cvt 313

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CVT-313 is a laboratory reagent developed by Santa Cruz Biotechnology. It is a selective inhibitor of the cell cycle regulator cyclin-dependent kinase 2 (CDK2). CVT-313 can be used in research applications to study cell cycle progression and regulation.

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Lab products found in correlation

Pen strep, by Thermo Fisher Scientific (2 mentions) Glutamax, by Thermo Fisher Scientific (2 mentions) Dulbecco modified eagle medium (dmem), by Corning (2 mentions) Palbociclib, by Selleck Chemicals (2 mentions) Stemdiffapel2, by Thermo Fisher Scientific (2 mentions) Recombinant human fgf basic fgf2 bfgf 146 aa protein, by R&D Systems (2 mentions) Pfhm 2, by Thermo Fisher Scientific (2 mentions) Stemdiffapel2, by STEMCELL (2 mentions) Recombinant human bmp 4 protein, by R&D Systems (2 mentions) Agn 194310, by MedChemExpress (2 mentions) Pd 0332991 isethionate, by Merck Group (2 mentions) Recombinant human vegf165, by Thermo Fisher Scientific (2 mentions) Pd0332991, by Selleck Chemicals (1 mentions) Cd19 mabs, by Southern Biotech (1 mentions) Jnk inhibitor 8, by Selleck Chemicals (1 mentions) Macsquant flow cytometer, by Miltenyi Biotec (1 mentions) Sp600125, by Selleck Chemicals (1 mentions) Sb203580, by Selleck Chemicals (1 mentions) Abt 737, by Selleck Chemicals (1 mentions) Aphidicolin, by Merck Group (1 mentions)

8 protocols using cvt 313

1

Foreskin Fibroblast Isolation and Treatment

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Fibroblasts were isolated from de-identified natal foreskin specimens and grown in DMEM (Cellgro) containing 15% fetal bovine serum supplemented with 1% Pen Strep (GIBCO), and 1% Glutamax (GIBCO), and incubated at 37°C with 5% CO2. Cells were treated with Palbociclib (PD-0332991) (SelleckChem # S1116) dissolved in sterile water or with CVT-313 (Santa Cruz Biotechnology, sc-221445) dissolved in DMSO by direct addition to media.
Schade A.E., Oser M.G., Nicholson H.E, & DeCaprio J.A. (2019). Cyclin D-CDK4 relieves cooperative repression of proliferation and cell cycle gene expression by DREAM and RB.. Oncogene, 38(25), 4962-4976.
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2

Directed Differentiation of hESCs into Cardiovascular Lineages

Cited 1 time
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H1-hESC or H9-Fucci-hESC were passaged using the method described above and re-plated onto Matrigel-coated plates. These hESC colonies were maintained in mTeSR for 24 hours, after which the cells were gently washed with DMEM:F12 and differentiated in chemically defined, serum-free, animal component-free medium STEMdiffAPEL2 (STEMCELL Technologies cat# 05270) supplemented with Protein-Free Hybridoma Medium (PFHM-II, GIBCO, cat# 12040077) by adding 5 mL of PFHM to 100 mL of STEMdiffAPEL2 medium. Growth factors depicted in Figure 1A were added sequentially (Stemolecule CHIR99021, 5 μM, Stemgent cat# 04–0004; Recombinant Human FGF basic/FGF2/bFGF (146 aa) Protein, 50 ng/ml, R&D systems cat# 233-FB-025; Recombinant Human BMP-4 Protein, 25 ng/ml, R&D systems cat# 314-BP-010; Recombinant Human VEGF165, 50 ng/ml, PeproTech, cat# 100–20) as described in Sriram et al. (2015) (link). For certain experiments, cells were treated with RA (0.5 μM, Sigma-Aldrich, cat# R2625–50MG), RAi (AGN 194310, 6nM, MedChemExpress, cat# HY-16681), CDK4/6i (PD 0332991 isethionate, 2nM, Sigma-Aldrich, cat# PZ0199–5MG), and/or CDK2i (CVT-313, 0.5 μM, Santa Cruz Biotechnology, cat# 199986–75-9) every 24hrs starting Day5.
Qiu J., Nordling S., Vasavada H.H., Butcher E.C, & Hirschi K.K. (2020). Retinoic Acid Promotes Endothelial Cell Cycle Early G1 State to Enable Human Hemogenic Endothelial Cell Specification. Cell reports, 33(9), 108465.
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3

Kinase Inhibition in Spleen Lysates

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Protein lysates from spleens collected from wild-type mice were incubated with 20 μM CVT-313 (Santa Cruz Biotechnology) for 30 min at room temperature. Subsequently, lysates were used for immunoprecipitation in the presence of 20 μM CVT-313 inhibitor. Washing buffer also contained 20 μM CVT-313.
Odajima J., Saini S., Jung P., Ndassa-Colday Y., Ficaro S., Geng Y., Marco E., Michowski W., Wang Y.E., DeCaprio J.A., Litovchick L., Marto J, & Sicinski P. (2016). Proteomic Landscape of Tissue-Specific Cyclin E Functions in Vivo. PLoS Genetics, 12(11), e1006429.
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4

Directed Differentiation of hESCs into Cardiovascular Lineages

Cited 1 time
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H1-hESC or H9-Fucci-hESC were passaged using the method described above and re-plated onto Matrigel-coated plates. These hESC colonies were maintained in mTeSR for 24 hours, after which the cells were gently washed with DMEM:F12 and differentiated in chemically defined, serum-free, animal component-free medium STEMdiffAPEL2 (STEMCELL Technologies cat# 05270) supplemented with Protein-Free Hybridoma Medium (PFHM-II, GIBCO, cat# 12040077) by adding 5 mL of PFHM to 100 mL of STEMdiffAPEL2 medium. Growth factors depicted in Figure 1A were added sequentially (Stemolecule CHIR99021, 5 μM, Stemgent cat# 04–0004; Recombinant Human FGF basic/FGF2/bFGF (146 aa) Protein, 50 ng/ml, R&D systems cat# 233-FB-025; Recombinant Human BMP-4 Protein, 25 ng/ml, R&D systems cat# 314-BP-010; Recombinant Human VEGF165, 50 ng/ml, PeproTech, cat# 100–20) as described in Sriram et al. (2015) (link). For certain experiments, cells were treated with RA (0.5 μM, Sigma-Aldrich, cat# R2625–50MG), RAi (AGN 194310, 6nM, MedChemExpress, cat# HY-16681), CDK4/6i (PD 0332991 isethionate, 2nM, Sigma-Aldrich, cat# PZ0199–5MG), and/or CDK2i (CVT-313, 0.5 μM, Santa Cruz Biotechnology, cat# 199986–75-9) every 24hrs starting Day5.
Qiu J., Nordling S., Vasavada H.H., Butcher E.C, & Hirschi K.K. (2020). Retinoic Acid Promotes Endothelial Cell Cycle Early G1 State to Enable Human Hemogenic Endothelial Cell Specification. Cell reports, 33(9), 108465.
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5

Measuring CLL Cell Apoptosis

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CLL cell apoptosis was measured in duplicates as previously described using the ApoScreen Annexin V Apoptosis Kit [29 (link)]. Briefly, cells were resuspended in 150 μl of Annexin V binding buffer containing 1 μl of Annexin V-PE, 1 μl of 7-aminoactinomycin D (7-AAD) and 1 μl of CD19-mAbs (Southern Biotech, Birmingham, AL) followed by flow cytometry on a MACSQuant flow cytometer (Miltenyi Biotec, San Diego, CA). Flow cytometry analysis was performed using FlowJo software (Tree Star, Ashland, OR). P1446A was provided by Piramal Healthcare Ltd. (Mumbai, India). JNK Inhibitor VIII, SP600125, SB203580, ABT-737 and PD0332991 were obtained from Selleck Chemicals (Houston, TX) and CVT-313—from Santa Cruz Biotechnology (Dallas, TX).
Paiva C., Godbersen J.C., Soderquist R.S., Rowland T., Kilmarx S., Spurgeon S.E., Brown J.R., Srinivasa S.P, & Danilov A.V. (2015). Cyclin-Dependent Kinase Inhibitor P1446A Induces Apoptosis in a JNK/p38 MAPK-Dependent Manner in Chronic Lymphocytic Leukemia B-Cells. PLoS ONE, 10(11), e0143685.
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6

Modulating Cell Signaling Dynamics

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For NCS treatment, medium was replaced with fresh medium supplemented with neocarzinostatin (N9162, Sigma‐Aldrich) during experiments. For myc overexpression, RPE cells were infected with fresh retrovirus containing MSCV‐Myc‐ER‐IRES‐GFP and 1 μl polybrene. Cells were subsequently passaged 48 of post‐infection and seeded onto a glass‐bottom plate for imaging. 16 h prior to imaging, tamoxifen was added at a final concentration of 50 nM. For aphidicolin treatment, medium was replaced with fresh medium supplemented with aphidicolin (A0781, Sigma‐Aldrich) for 8 h during experiments, washed off once with PBS, and then replenished with imaging media described below. For CDK2 inhibition, cells were treated with 2 μM CVT‐313 (221445, Santa Cruz) prior to starting the imaging.
Chao H.X., Fakhreddin R.I., Shimerov H.K., Kedziora K.M., Kumar R.J., Perez J., Limas J.C., Grant G.D., Cook J.G., Gupta G.P, & Purvis J.E. (2019). Evidence that the human cell cycle is a series of uncoupled, memoryless phases. Molecular Systems Biology, 15(3), e8604.
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7

CVT-313 Effects on Cell Culture

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Cells were cultured in the presence of 10 or 20 μM CVT-313 (Santa Cruz) for 48 hr.
Liu L., Michowski W., Inuzuka H., Shimizu K., Nihira N.T., Chick J.M., Li N., Geng Y., Meng A.Y., Ordureau A., Kolodziejczyk A., Ligon K.L., Bronson R.T., Polyak K., Harper J.W., Gygi S.P., Wei W, & Sicinski P. (2017). G1 cyclins link proliferation, pluripotency and differentiation of embryonic stem cells. Nature cell biology, 19(3), 177-188.
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8

Foreskin Fibroblast Isolation and Treatment

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Fibroblasts were isolated from de-identified natal foreskin specimens and grown in DMEM (Cellgro) containing 15% fetal bovine serum supplemented with 1% Pen Strep (GIBCO), and 1% Glutamax (GIBCO), and incubated at 37°C with 5% CO2. Cells were treated with Palbociclib (PD-0332991) (SelleckChem # S1116) dissolved in sterile water or with CVT-313 (Santa Cruz Biotechnology, sc-221445) dissolved in DMSO by direct addition to media.
Schade A.E., Oser M.G., Nicholson H.E, & DeCaprio J.A. (2019). Cyclin D-CDK4 relieves cooperative repression of proliferation and cell cycle gene expression by DREAM and RB.. Oncogene, 38(25), 4962-4976.
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