The chemical structure of imatinib (Sigma, with a purity ≥98%) is shown in Figure 1 . The colon cancer cell line SW480 was provided by the Shanghai Cell Bank of the Chinese Academy of Sciences. Fetal bovine serum (FBS) was purchased from Hyclone, Leibowitz’s L-15 medium and puromycin were bought from Gibco company, and rabbit anti-GAPDH, anti-HGF, anti-p21, and anti-p27 were purchased from Sigma-Aldrich.
Anti p27
Manufactured by Merck Group
Sourced in United Kingdom
Anti-p27 is a laboratory reagent used for the detection and quantification of the p27 protein. The p27 protein is a cyclin-dependent kinase inhibitor that plays a role in cell cycle regulation. The Anti-p27 reagent can be used in various analytical techniques, such as Western blotting and immunohistochemistry, to measure the expression levels of p27 in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti p53, by Merck Group (2 mentions)
Amersham ecl select western blotting detection reagent, by GE Healthcare (2 mentions)
Anti bax, by Abcam (2 mentions)
Imatinib, by Merck Group (1 mentions)
Puromycin, by Thermo Fisher Scientific (1 mentions)
Anti p21, by Merck Group (1 mentions)
Leibowitz s l15 medium, by Thermo Fisher Scientific (1 mentions)
Rabbit anti gapdh, by Merck Group (1 mentions)
Hrp conjugated goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
Hrp conjugated goat anti mouse igg, by Thermo Fisher Scientific (1 mentions)
Caspase 3, by Cell Signaling Technology (1 mentions)
Anti mmp2, by Santa Cruz Biotechnology (1 mentions)
Anti vegf, by Abcam (1 mentions)
Anti β actin, by Cell Signaling Technology (1 mentions)
Anti bcl 2, by Abcam (1 mentions)
Anti cyclin d1, by Merck Group (1 mentions)
Anti cdk4, by Merck Group (1 mentions)
Histoclear, by National Diagnostics (1 mentions)
Anti cdk2, by Santa Cruz Biotechnology (1 mentions)
Anti cyclin d1, by Cell Signaling Technology (1 mentions)
4 protocols using anti p27
1
Imatinib Effects on SW480 Colon Cancer Cells
2
Western Blot Analysis of Tumor Markers
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Total proteins prepared from the tumor tissue were separated by 4–20% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) for 2 h, after which the resolved proteins were transferred to nitrocellulose membranes for 2 h at 40 V. Each membrane was then incubated separately, overnight at 4°C, with the following primary antibodies: anti-p53 (Sigma-Aldrich Co.), anti-p27 (Sigma-Aldrich Co.), anti-bax (Abcam, Cambridge, UK), anti-Bcl2 (Abcam), caspase-3 (Cell Signaling Technology, Danvers, MA, USA), anti-MMP2 (Santa Cruz Biotechnology, Inc. Santa Cruz, CA, USA), anti-VEGF (Abcam), and anti-β-actin (Cell Signaling Technology). The membranes were then washed with washing buffer (137 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, and 0.05% Tween 20) and incubated with HRP-conjugated goat anti-rabbit IgG (Invitrogen) and HRP-conjugated goat anti-mouse IgG (Invitrogen) at a 1:1,000 dilution, at room temperature for 1 h. Membrane blots were developed using Amersham ECL Select Western Blotting detection reagent (GE Healthcare Piscataway, NJ, USA).
3
Immunohistochemical analysis of cell cycle proteins
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Embryos were fixed for 24 h in cold 4% paraformaldehyde in PBS (PFA), washed x 3 in PBS, dehydrated, cleared in Histoclear (National Diagnostics), embedded in paraffin wax and sectioned at 8 μm thickness. Sections were dewaxed, microwaved in 0.1 M citric acid buffer, washed in PBS-Triton 0.3% (PBS-T) for 10 min, then treated with 3% hydrogen peroxide. Sections were blocked in PBS-T containing 5% FCS and 5% normal goat serum for 2 h. Primary antibodies were applied overnight at 4 °C: anti-Cdk4 (Sigma, C8218; 1:80 dilution), anti-Cyclin D1 (Sigma, C7464; 1:40 dilution) and anti-p27 (Sigma, P2092; 1:80 dilution). Secondary antibody was biotinylated rabbit anti-mouse IgG (1:400 dilution), detected using Avidin-Biotin-Complex/horseradish peroxidase (DAKO) and diaminobenzidine. Three embryos at each Mode of neurulation were assessed with each antibody.
4
Western Blot Analysis of Skin Proteins
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Proteins prepared from the skin tissue of mice were separated by 4–20% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) for 2 h, after which the resolved proteins were transferred to nitrocellulose membranes for 2 h at 40 V. Each membrane was then incubated separately, overnight at 4℃, with the following primary antibodies: anti-p27 (1:1,000, Sigma-Aldrich Co.), anti-p53 (1:1,000, Sigma-Aldrich Co.), anti-Bax (1:1,000, Abcam, Cambridge, UK), anti-Bcl-2 (1:1,000, Thermo Fisher Scientific, Waltham, MA, USA), anti-Caspase-3 (1:1,000, Cell Signaling Technology, Danvers, MA, USA), anti-Cyclin D1 (1:1,000, Cell Signaling Technology), anti-CDK2 (1:1,000, Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-CDK4 (1:1,000, Santa Cruz Biotechnology) and anti-β-actin (1:1,000, Sigma-Aldrich Co.). The membranes were subsequently washed with washing buffer (137 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, and 0.05% Tween 20) and incubated with horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (1:2,000 dilution, Thermo Fisher Scientific) at room temperature for 1 h. Membrane blots were developed using Amersham ECL Select Western Blotting detection reagent (GE Healthcare, Little Chalfont, UK). The density of each band was quantified using the Image Analyzer System (Fluorchem FC2, Alpha Innotech, CA, USA) and was expressed as a fold-increase over control values.
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