Protein lysate (PMSF: RIPA=1:100) (Solarbio) was used to lyse the heart tissue from the infarct marginal zone. The extracted proteins were subjected to polyacrylamide gel electrophoresis (GenScript, Nanjing, China) and transferred onto polyvinylidene di uoride membranes (Thermo Fisher Scienti c). The membranes were blocked with QuickBlock Western reagent (Beyotime, Beijing, China) and incubated sequentially with CCR2 overnight at 4°C. Subsequently, the washed membranes were incubated with the corresponding HRP-conjugated secondary antibodies (SA00001-2; Proteintech) for 2 h at 25℃. Lastly, a gel imaging system (GE Healthcare, CHI, USA) was used to quantify the protein bands. The western blots were quanti ed using ImageJ software (NIH, MD, USA).
Polyacrylamide gel electrophoresis
Manufactured by GenScript
Sourced in China
Polyacrylamide gel electrophoresis (PAGE) is a laboratory technique used to separate and analyze biomolecules, such as proteins or nucleic acids, based on their size and charge. The process involves applying an electric field to a polyacrylamide gel, which causes the biomolecules to migrate through the gel at different rates, allowing for their separation and identification.
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2 protocols using polyacrylamide gel electrophoresis
1
Quantitative Western Blotting of CCR2
2
Quantitative Western Blotting of CCR2
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Protein lysate (PMSF: RIPA=1:100) (Solarbio) was used to lyse the heart tissue from the infarct marginal zone. The extracted proteins were subjected to polyacrylamide gel electrophoresis (GenScript, Nanjing, China) and transferred onto polyvinylidene di uoride membranes (Thermo Fisher Scienti c). The membranes were blocked with QuickBlock Western reagent (Beyotime, Beijing, China) and incubated sequentially with CCR2 overnight at 4°C. Subsequently, the washed membranes were incubated with the corresponding HRP-conjugated secondary antibodies (SA00001-2; Proteintech) for 2 h at 25℃. Lastly, a gel imaging system (GE Healthcare, CHI, USA) was used to quantify the protein bands. The western blots were quanti ed using ImageJ software (NIH, MD, USA).
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