Total protein of mouse hindlimbs and BMSCs was extracted using the RIPA lysis buffer (CWBIOTECH, China) containing proteinase and phosphatase inhibitors (CWBIOTECH, China), and then quantified by the BCA method (FudeBio, China). The proteins were separated by SDS-PAGE gel electrophoresis (Smart-Lifesciences, China) and transferred to a polyvinylidene fluoride (PVDF) membrane (Millipore, USA). After 1 h blocking by 5% BSA, membranes were incubated at 4 °C overnight with relevant primary antibodies. The PVDF membranes were incubated in HRP conjugated secondary antibody for 1 h at room temperature. Immunoreactivities were detected by a chemiluminescence kit (FudeBio, China). Data were normalized to GAPDH and quantified by ImageJ. The antibodies used in this study were listed in Table 2 .
Proteinase and phosphatase inhibitors
Manufactured by CWBIO
Sourced in China
Proteinase and phosphatase inhibitors are laboratory reagents used to prevent the breakdown of proteins and the dephosphorylation of phosphorylated proteins during sample preparation and analysis. They are commonly used in biochemical and cell biology research to preserve the integrity of proteins and their post-translational modifications.
Automatically generated - may contain errors
3 protocols using proteinase and phosphatase inhibitors
1
Western Blot Analysis of Protein Expression
2
Western Blot Analysis of Protein Levels
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Total protein of mouse hindlimbs and BMSCs was extracted using the RIPA lysis buffer (CWBIOTECH, China) containing proteinase and phosphatase inhibitors (CWBIOTECH, China), and then quanti ed by the BCA method (FudeBio, China). The proteins were separated by SDS-PAGE gel electrophoresis (Smart-Lifesciences, China) and transferred to a polyvinylidene uoride (PVDF) membrane (Millipore, USA). After 1h blocking by 5% BSA, membranes were incubated at 4°C overnight with relevant primary antibodies.
The PVDF membranes were incubated in HRP conjugated secondary antibody for 1 h at room temperature. Immunoreactivities were detected by a chemiluminescence kit (FudeBio, China). Data were normalized to GAPDH and quanti ed by ImageJ. The antibodies used in this study were listed in Table 2.
The PVDF membranes were incubated in HRP conjugated secondary antibody for 1 h at room temperature. Immunoreactivities were detected by a chemiluminescence kit (FudeBio, China). Data were normalized to GAPDH and quanti ed by ImageJ. The antibodies used in this study were listed in Table 2.
3
Western Blot Analysis of Protein Levels
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total protein of mouse hindlimbs and BMSCs was extracted using the RIPA lysis buffer (CWBIOTECH, China) containing proteinase and phosphatase inhibitors (CWBIOTECH, China), and then quanti ed by the BCA method (FudeBio, China). The proteins were separated by SDS-PAGE gel electrophoresis (Smart-Lifesciences, China) and transferred to a polyvinylidene uoride (PVDF) membrane (Millipore, USA). After 1h blocking by 5% BSA, membranes were incubated at 4°C overnight with relevant primary antibodies.
The PVDF membranes were incubated in HRP conjugated secondary antibody for 1 h at room temperature. Immunoreactivities were detected by a chemiluminescence kit (FudeBio, China). Data were normalized to GAPDH and quanti ed by ImageJ. The antibodies used in this study were listed in Table 2.
The PVDF membranes were incubated in HRP conjugated secondary antibody for 1 h at room temperature. Immunoreactivities were detected by a chemiluminescence kit (FudeBio, China). Data were normalized to GAPDH and quanti ed by ImageJ. The antibodies used in this study were listed in Table 2.
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