The genotypes of p53 mice were identified by PCR. DNA was extracted from mice via the tail. DreamTaq Green PCR Master Mix for PCR was purchased from Thermo Fisher Scientific, Inc. (cat. no. K1081). The wild-type primers used were as follows: Wild-type primer 1, 5'-CAGCGTGGTGGTACCTTAT-3'; and wild-type primer 2, 5'-CTATCAGGACATAGCGTTGG-3', with a PCR product size of 450 bp. The mutant primers used were as follows: Mutation primer 3, 5'-TATACTCAGAGCCGGCCT-3'; and mutation primer 4, 5'-CTATCAGGACATAGCGTTGG-3', with a PCR product size of 615 bp. PCR conditions were as follows: Denaturation at 94˚C for 3 min, 94˚C for 1 min, 60˚C for 2 min and 72˚C for 2 min, for 30 cycles, followed by extension at 72˚C for 5 min. Gel electrophoresis was performed using 1.2% agarose gel. Images were captured with an automatic gel imaging system (Syngene).
Automatic gel imaging system
Manufactured by Syngene
The Automatic Gel Imaging System is a laboratory equipment designed to capture and analyze digital images of electrophoresis gels, such as those used in DNA, RNA, or protein analysis. The system automatically photographs the gel and provides digital image files for further processing and evaluation.
Automatically generated - may contain errors
Lab products found in correlation
Sqstm1 p62, by Abcam (1 mentions)
Bnip3l nix, by Cell Signaling Technology (1 mentions)
Phospho ampkα, by Cell Signaling Technology (1 mentions)
Sglt2, by Abcam (1 mentions)
Ecl system, by Advansta (1 mentions)
Phospho mtor, by Cell Signaling Technology (1 mentions)
Ampkα, by Cell Signaling Technology (1 mentions)
2 protocols using automatic gel imaging system
1
Genotyping p53 Mice by PCR
2
Protein Expression Analysis in Renal Tissues
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Renal tissues and cells were homogenized in RIPA buffer containing Protease/Phosphatase Inhibitor Cocktail (Beijing Suo Lai Bao Technology Co., Ltd.) after washing with PBS, and total protein concentration was estimated using BCA Protein Assay Kit (Wuhan Doctorate Bioengineering Co., Ltd.). Protein samples (40-80μg) were submitted to SDS-PAGE and then transferred to nitrocellulose membranes. Membranes were blocked for 2 h in 5% non-fat milk and then incubated with primary antibodies against LC3 (1:1,000, Cell Signaling), SQSTM1/p62 (1:1,000, Abcam), BNIP3L/Nix (1:1,000, Cell Signaling), mTOR (1:1,000, Cell Signaling), Phospho-mTOR (1:1,000, Cell Signaling), AMPKα (1:1,000, Cell Signaling), Phospho-AMPKα (1:1,000, Cell Signaling), SGLT2 (1:1,000, Abcam), and β-actin (1:500, Sanjian Biotechnology) at 4°C overnight. After being washed with TBST, the membranes were incubated with peroxidase-conjugated secondary antibodies (1:5000, Ai Meijie Technology Co., Ltd, China). The reactive bands were detected using the ECL system (Advansta). Signal intensity was then assessed using automatic gel imaging system (SYNGENE). Studies were replicated 3 times.
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