Anti cdk2

Manufactured by Bioworld Technology

Sourced in United States

Anti-CDK2 is a laboratory reagent that functions as an inhibitor of the cyclin-dependent kinase 2 (CDK2) protein. CDK2 is a critical regulator of cell cycle progression, and inhibition of its activity can have implications for research in various fields, including cell biology and oncology.

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4 protocols using anti cdk2

Comprehensive Protein Analysis Protocol

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IU1, siRNA and shRNA were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). MG132 and bortezomib (Velcade) were purchased from Selleckchem (Houston, TX, USA). MTS assay (CellTiter 96 Aqueous One Solution reagent) was purchased from Promega Corporation (Madison, WI, USA). Propidium iodide (PI) and Annexin V-FITC Apoptosis Detection Kit were purchased from Keygen Company (Nanjing, China). Dynabeads antibody coupling kit was from Life technologies. Antibodies used in this study were purchased from following sources: anti-ubiquitin (P4D1) (Santa Cruz Biotechnology); anti-PARP, anti-CDK2, anti-phospho-Rb, anti-Rb, anti-PSA, anti-Bax, anti-GFP, anti-GAPDH (Bioworld Technology, Inc., Louis Park, MN, USA); anti-CDK4, anti-CDK6, anti-phospho-MDM2, anti-P53, anti-USP14, anti-Flag, anti-cyclin D1, anti-p15, anti-p27 (Cell Signaling Technology, Beverly, MA, USA); anti-MDM2 and anti-AR (Abcam, USA).

Liao Y., Liu N., Hua X., Cai J., Xia X., Wang X., Huang H, & Liu J. (2017). Proteasome-associated deubiquitinase ubiquitin-specific protease 14 regulates prostate cancer proliferation by deubiquitinating and stabilizing androgen receptor. Cell Death & Disease, 8(2), e2585-.

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Molecular Mechanisms Underlying Autophagy Regulation

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Spautin-1(S7888), 3-methyladanine (3-MA, S2767), Z-VAD-FMK (S7023), SP600125 (S1460), SB230580 (S1076), LY3214996 (S8534), Gefitinib (S1025) and Enzalutamide (MDV3100, S1250) were purchased from Sellectchem (Houston, TX, USA). SKP2-C25 (M60136) was obtained from Xcessbio Biosciences, Inc. (San Diego, CA). USP10 (sc-76,811), USP13 (sc-76,815) and Glut1 (sc-35,493) siRNAs were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies: anti-Glut1 (ab652), anti-USP13 (ab109264) (Abcam, Cambridge, MA); anti-GAPDH (BS60630), anti-Ki67 (BS1454) (Bioworld Technology, Inc., Louis Park, MN, USA); anti-USP10(#8501), anti-SKP2(#2652), anti-p27(#3686), anti-CDK4(#12790), anti-CDK2(#2546), anti-CyclinD1(#2978), anti-p15(#4822), anti-p21(#2947), anti-PARP(#9532), anti-Bim(#2933), anti-Bax(#14796), anti-Bcl-2(#15071), anti-activated Caspase-3(#9664), anti-phospho-JNK (#9255), anti-JNK(#9252), anti-phospho-ERK1/2(#4370), anti-ERK1/2(#4695), anti-phospho-p38(#4511), anti-p38(#8690), anti-phospho-MKK4(#4514), anti-MKK4(#9152), anti-phospho-MEK1/2 (#9154), anti-MEK1/2(#4694), anti-phospho-EGFR (Tyr1173)(#4407), anti-phospho-EGFR (Tyr1068)(#3777), anti-EGFR(#2085) (Cell Signaling Technology, Beverly, MA, USA).

Liao Y., Guo Z., Xia X., Liu Y., Huang C., Jiang L., Wang X., Liu J, & Huang H. (2019). Inhibition of EGFR signaling with Spautin-1 represents a novel therapeutics for prostate cancer. Journal of Experimental & Clinical Cancer Research : CR, 38, 157.

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Comprehensive Protein Expression Analysis

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Total proteins were isolated from cells or tissues using the RIPA lysis buffer (Beyotime, China) and subjected to 10% polyacrylamide gel electrophoresis. After transferring to PVDF membrane, blocking with 5% skim milk, incubation with primary antibodies and secondary antibodies, the proteins on the membrane were finally detected by ECL kit (EpiZyme, China) and imaging analysis system (BioRad, USA). The primary antibodies used in this study were as follows: anti-GAPDH (Bioworld, 1:10000), anti-CDK2 (Bioworld, 1:1000), anti-Cyclin E1 (Bioworld, 1:1000), anti-CDK4 (Bioworld, 1:1000), anti-Cyclin D1 (Bioworld, 1:1000), anti-p53 (Bioss, 1:500), anti-p21 (Bioworld, 1:500), anti-p16 (Bioss, 1:1000), anti-ubiquitin (Santa, 1:500), anti-FTO (Bioss, 1:1000), anti-YTHDF2 (Abcam, 1:1000).

Cui Y., Zhang C., Ma S., Li Z., Wang W., Li Y., Ma Y., Fang J., Wang Y., Cao W, & Guan F. (2021). RNA m6A demethylase FTO-mediated epigenetic up-regulation of LINC00022 promotes tumorigenesis in esophageal squamous cell carcinoma. Journal of Experimental & Clinical Cancer Research : CR, 40, 294.

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Western Blotting of Cellular Proteins

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We used 10% SDS-PAGE to dilute the corresponding proportion of protein lysates (1:4) for electrophoresis and transferred the proteins to PVDF membranes (Millipore, MA) with cold transfer buffer. The membrane was sealed with 5% skim milk in TBS-T for 1 hour at room temperature (RT) and then incubated with the relevant primary antibodies overnight at 4°C. After washing the membranes, we subsequently incubated them with secondary antibodies bound to horseradish peroxidase (HRP). Enhanced chemiluminescence was used to visualize the proteins. Western blotting was performed using the following antibodies: anti-actin (Sigma-Aldrich, 1:8,000) and anti-CDK2 (Bioworld, MN). We subsequently used goat anti-mouse IgG (HRP) and goat anti-rabbit IgG (HRP) diluted at 1:2000 as secondary antibodies.

Liu Y., Tang J., & Dandan Y. (2022). MiR-362-5p targets CDK2 and inhibits tumorigenesis in renal cell carcinoma.

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