Total protein samples were extracted from cultured BMSCs using the Total Protein Extraction kit (#C006225-0050; Sangon Biotech, Shanghai, China) according to the manufacturer’s instructions. The protein concentrations were determined by the bicinchoninic acid method. Approximately 30 μg of protein was then boiled at 100 °C for 5 min, separated by 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis, and blotted onto polyvinylidene fluoride (PVDF) membranes The membranes were subsequently blocked in 5% lipid-free milk solution, incubated with diluted primary antibodies, washed three times with TBST solution, and incubated with diluted secondary antibodies. Finally, the PVDF membranes were developed with the EasyBlot ECL (enhanced chemiluminescence) kit (#D601039-0050; Sangon Biotech, Shanghai, China) according to the manufacturer’s instructions. At least three biological replicates were conducted for protein quantitation, and GAPDH was used as an internal standard. The antibodies used in the present study included anti-ALP (#ab229126; Abcam), anti-osteopontin (OPN) (#ab8448; Abcam), anti-runt-related transcription factor 2 (RUNX2) (#12556; CST), anti-GAPDH (#ab181602; Abcam), and anti-growth arrest-specific gene 1 (GAS1) (#ab236618; Abcam).
Easyblot ecl
Manufactured by Sangon
The EasyBlot ECL is a chemiluminescent western blotting detection system designed for sensitive and reliable protein detection. It provides a simple and efficient method for visualizing proteins on western blots.
Automatically generated - may contain errors
3 protocols using easyblot ecl
1
Western Blot Analysis of Osteogenic Markers
2
Protein Extraction and Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Total protein samples were extracted from cultured BMSCs using the Total Protein Extraction kit (#C006225-0050; Sangon Biotech, Shanghai, China) according to the manufacturer's instructions. The protein concentrations were determined by the bicinchoninic acid method. Approximately 30 µg of protein was then boiled at 100°C for 5 min, separated by 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis, and blotted onto polyvinylidene uoride (PVDF) membranes The membranes were subsequently blocked in 5% lipidfree milk solution, incubated with diluted primary antibodies, washed three times with TBST solution, and incubated with diluted secondary antibodies. Finally, the PVDF membranes were developed with the EasyBlot ECL (enhanced chemiluminescence) kit (#D601039-0050; Sangon Biotech, Shanghai, China) according to the manufacturer's instructions. At least three biological replicates were conducted for protein quantitation, and GAPDH was used as an internal standard. The antibodies used in the present study included anti-ALP (#ab229126; Abcam), anti-osteopontin (OPN) (#ab8448; Abcam), anti-runt-related transcription factor 2 (RUNX2) (#12556; CST), anti-GAPDH (#ab181602; Abcam), and anti-growth arrest-speci c gene 1 (GAS1) (#ab236618; Abcam).
3
Protein Extraction and Western Blot Analysis of BMSC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total protein samples were extracted from cultured BMSCs using the Total Protein Extraction kit (#C006225-0050; Sangon Biotech, Shanghai, China) according to the manufacturer's instructions. The protein concentrations were determined by the bicinchoninic acid method. Approximately 30 µg of protein was then boiled at 100 °C for 5 min, separated by 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis, and blotted onto polyvinylidene uoride (PVDF) membranes The membranes were subsequently blocked in 5% lipid-free milk solution, incubated with diluted primary antibodies, washed three times with TBST solution, and incubated with diluted secondary antibodies. Finally, the PVDF membranes were developed with the EasyBlot ECL (enhanced chemiluminescence) kit (#D601039-0050; Sangon Biotech, Shanghai, China) according to the manufacturer's instructions. At least three biological replicates were conducted for protein quantitation, and GAPDH was used as an internal standard. The antibodies used in the present study included anti-ALP (#ab229126; Abcam), anti-osteopontin (OPN) (#ab8448; Abcam), anti-runt-related transcription factor 2 (RUNX2) (#12556; CST), anti-GAPDH (#ab181602; Abcam), and anti-growth arrest-speci c gene 1 (GAS1) (#ab236618; Abcam).
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