Cells were collected and lysed with cell lysis buffer for western blotting (Beyotime, Haimen, China). The proteins (30 μg per lane) were separated on 12% SDS-polyacrylamide gels and transferred on a polyvinylidene fluoride membrane (Millipore, Billerica, MA, USA). Immunoblotting of the membrane was performed using the following primary antibodies: anti-CDK6 (Boster, Wuhan, China), anti-cyclin D1 (Affinity, Cincinnati, OH, USA), anti-MMP-2 (Boster), anti-MMP-9 (Boster) and anti-β-actin (4 A Biotech, Beijing, China). The signals were revealed after incubation with the recommended secondary antibodies using an Odyssey Infrared Imaging System (LI-COR, Lincoln, NE, USA). β-actin was used as the control.
Anti cdk6
Manufactured by Boster Bio
Sourced in China
Anti-CDK6 is a laboratory reagent that functions as an inhibitor of the Cyclin-Dependent Kinase 6 (CDK6) protein. CDK6 is a key regulator of the cell cycle and plays a crucial role in cell division and proliferation. Anti-CDK6 can be utilized in experimental settings to study the role of CDK6 in various cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Anti cdk4, by Boster Bio (2 mentions)
Anti β actin, by 4A Biotech (1 mentions)
Anti cyclin d1, by Affinity Biosciences (1 mentions)
Polyvinylidene fluoride membrane, by Merck Group (1 mentions)
Odyssey infrared imaging system, by LI COR (1 mentions)
Cell lysis buffer for western blotting, by Beyotime (1 mentions)
Anti mmp2, by Boster Bio (1 mentions)
Anti mmp 9, by Boster Bio (1 mentions)
Anti akt, by Cell Signaling Technology (1 mentions)
Pictilisib, by Selleck Chemicals (1 mentions)
Anti foxo3a, by Cell Signaling Technology (1 mentions)
Anti cleaved caspase 3, by Beyotime (1 mentions)
Anti fasl, by Cell Signaling Technology (1 mentions)
Anti trail, by Cell Signaling Technology (1 mentions)
Anti parp, by Beyotime (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Sartorius (1 mentions)
Anti p akt, by Abcam (1 mentions)
Anti erk, by Abcam (1 mentions)
Sorafenib, by Selleck Chemicals (1 mentions)
3 protocols using anti cdk6
1
Western Blot Analysis of Cell Signaling
2
Sorafenib and FOXO3a Regulation in Apoptosis
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PD was purchased from Must Bio-Technology Co., Ltd. (Chengdu, China) and sorafenib was purchased from Selleckchem (Houston, TX, USA). Fetal bovine serum (FBS) was purchased from Bioind (Biological Industries, BeitHaEmek, Israel). The anti-FOXO3a, anti-FasL, anti-Bim, anti-TRAIL, anti-Akt and anti-Ubiquitin antibodies were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). The anti-p-Akt, anti-ERK and anti-p-ERK antibodies were obtained from Abcam (Shanghai, China). The anti-Caspase 3, anti-Cleaved Caspase 3, anti-Caspase 3 and anti-PARP antibodies were obtained from Beyotime Biotechnology, Inc. (Shanghai, China). The anti-CDK4, anti-CDK6 and anti-cyclin D antibodies were obtained from Boster Biological Technology, Inc. (Wuhan, China). The Z-VAD-FMK, MG132, pictilisib and MK2206 were purchased from Selleckchem (Houston, TX, USA). Lipofectamine™ 2000 was purchased from Invitrogen (Shanghai, China). The shRNA-FOXO3a plasmid, cDNA-Akt plasmid and cDNA-FOXO3a were purchased Shanghai GeneChem Co., Ltd (Shanghai, China).
3
Protein Extraction and Western Blot Analysis
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Proteins were extracted using NP40 buffer (Beyotime Institute of Biotechnology, China) supplemented with protease inhibitor phenylmethanesulfonyl fluoride (PMSF; Roche, Switzerland). The protein concentrations were calculated using the bicinchoninic acid (BCA) protein assay kit (Beyotime Institute of Biotechnology, China). Equivalent amounts of 30 µg protein samples were isolated using 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Then, western blot analysis was performed following a standard procedure with the primary antibodies: anti-p21 (1:1000; Cell Signaling Technology, USA), anti-cyclin D1 (1:500; Boster, China), anti-CDK4 (1:500; Boster), anti-CDK6 (1:500; Boster), and anti-E-cadherin (1:1000; Cell Signaling Technology). The anti-β-Actin (Boster) was used as an internal control at 1:500 dilution. And then, protein bands were incubated with secondary antibodies (1:5000) conjugated with horseradish peroxidase (HRP) and visualized using the enhanced chemiluminescence (ECL) assay kit.
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