The production of MDA, a marker of lipid peroxidation, was determined to assess oxidative injury using a commercial MDA assay kit (Nanjing, China). Brie y, H9C2 cells were seeded in 6-well plates for 24 h prior to various treatments. Following treatment, H9C2 cells were washed with ice-cold PBS and incubated with RIPA lysis buffer (Nanjing, China) for 10 min at 4°C. The lysates were centrifuged at 12,000 rpm for 10 min at 4°C, and the supernatant were collected for detection of MDA according to the manufacturer's instructions. The determination of SOD (Nanjing, China) is the same as above. All operations were carried out following the manufacture's instructions.
Ripa lysis buffer
Manufactured by Nanjing Chemical Reagent
Sourced in China
RIPA lysis buffer is a widely used detergent-based cell lysis and extraction buffer. It is designed to solubilize cellular components, including proteins, for further analysis.
Automatically generated - may contain errors
Lab products found in correlation
Mda assay kit, by Nanjing Chemical Reagent (1 mentions)
Gapdh polyclonal antibody, by Proteintech (1 mentions)
Interleukin 6 (il 6), by Cell Signaling Technology (1 mentions)
Cd206, by Abcam (1 mentions)
Stat3, by Cell Signaling Technology (1 mentions)
Phospho stat3, by Cell Signaling Technology (1 mentions)
2 protocols using ripa lysis buffer
1
Lipid Peroxidation and Antioxidant Assessment
2
Protein Expression Analysis in Myocardial Tissue
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Fresh myocardial tissue was lysed by RIPA lysis buffer (Nanjing, China) and proteins were extracted. Proteins were separated by SDS-PAGE and transferred to PVDF membranes. The membrane was closed with 5% bovine serum albumin and then incubated with the appropriate primary antibody overnight at 4 °C, followed by washing with TBS-Tween and incubation with the appropriate secondary antibody at room temperature for 1 h. Spots were detected using an ECL system (Amersham, UK) and optical density was measured using ImageJ software. The following antibodies were used: iNOS (Cell Signaling Technology, 13120S), CD206 (Abcam, ab125028), (Abcam, ab28946), IL-6 (Cell Signaling Technology, 12912S), Stat3 (Cell Signaling Technology, 9139S), Phospho-Stat3 (Cell Signaling Technology, 9145S), GAPDH polyclonal antibody (Proteintech, No.10494–1-AP).
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