Total protein from MCF-7 breast cancer cells was extracted after 6-TG treatment. The protein samples were fractionated on a BiofurawTM Precast Gel (Tanon, China) and electrically transferred onto a polyvinylidene difluoride (PVDF) membrane. Next, the membranes were blocked with 5% non-fat milk and incubated overnight at 4°C with a primary antibody (DNMT1 antibody from GeneTex, USA; p53 antibody from Affinity, USA; FAS/CD95anyibody from Proteintech, China). The membranes were washed 3 times with TBST and incubated for 1–2 h at room temperature (RT) with a secondary antibody (horseradish peroxidase-conjugated goat anti-rabbit IgG and goat anti-mouse IgG from Proteintech, China). Finally, the membranes were washed 3 times with TBST and visualized by using a Tanon 5200 automatic fluorescence/chemiluminescence imaging analysis system (Tanon, China).
P53 antibody
Manufactured by Affinity Biosciences
Sourced in United States, China
The P53 antibody is a laboratory reagent used to detect the presence of the p53 protein in biological samples. P53 is a tumor suppressor protein that plays a crucial role in regulating cell growth and division. The antibody is designed to specifically bind to and identify the p53 protein, allowing researchers to study its expression and function in various cellular and molecular processes.
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Lab products found in correlation
2 protocols using p53 antibody
1
Protein Expression Analysis in MCF-7 Cells
2
Western Blot Analysis of Cell Signaling Proteins
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Cells were collected in RIPA lysis buffer, and the proteins were quantified using a BCA Protein Concentration Assay Kit (MDL Biotechnology, China). The protein was isolated on a 10 % SDS-PAGE gel (MDL Biotechnology, China), and transferred onto PVDF membranes. Then, the membranes were blocked with 5 % (w/v) nonfat dry milk in PBS and 0.05 % Tween. The primary antibodies used as follows: anti-FAK antibody (1:1,000; MDL Biotechnology, China), anti-CDC27 antibody (1:1,000; MDL Biotechnology, China), RPRD1A-antibody (1:1,000; MDL Biotechnology, China), RPL34-antibody (1:1,000; MDL Biotechnology, China), RPL24-antibody (1:1,000; MDL Biotechnology, China), EIF6-antibody (1:1,000; MDL Biotechnology, China), p53-antibody (1:1000; Affinity, USA), MDM2-antibody (1:1000; Abcam, USA), p21-antibody (1:1000; Epitmics, USA), CDK2-antibody (1:3000; Abcam, USA), CDK4-antibody (1:3000; Abcam, USA), RB-antibody (1:2000; Abcam, USA), ATM-antibody (1:3000; Abcam, USA), E2F1-antibody (1:1000; Abcam, USA). β-actin (1:1000; MDL Biotechnology, China) or GAPDH (1:5000; Abcam, USA) was used as the loading control. The secondary antibodies were incubated with goat IgG-HRP (1:5,000; MDL Biotechnology, China) at room temperature for one hour. The blots were detected using a ChemiDoc MP chemiluminescence imaging system (Bio-rad, USA).
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