The protein was extracted from granulosa cells using RIPA lysis buffer (Beyotime, China) with proteinase and phosphatase inhibitors. The protein concentration was measured by a BCA protein assay kit (Beyotime, China). The samples were denatured at 95 °C for 10 min with 5× protein-loading buffer. The protein was separated by 10% SDS-PAGE and then was transferred onto 0.22-μm PVDF membranes (Millipore, USA). Next, the membranes were blocked with 5% skim milk for 1 h and were then incubated with primary antibodies at 4 °C overnight. Subsequently, the membranes were washed three times and incubated with HRP-conjugated goat anti-rabbit or goat anti-mouse secondary antibodies (1:3000, Abcam, USA) for 1 h. Finally, the proteins were detected by enhanced chemiluminescence and were imaged by an Image Quant LAS 4000 mini biomolecular imager (Bio-Rad, USA).
Hrp conjugated goat anti rabbit or goat anti mouse secondary antibodies
Manufactured by Abcam
Sourced in United States
HRP-conjugated goat anti-rabbit or goat anti-mouse secondary antibodies are laboratory reagents used to detect the presence of primary antibodies specific to rabbit or mouse proteins in various immunoassay techniques, such as Western blotting, ELISA, and immunohistochemistry. These secondary antibodies are conjugated with horseradish peroxidase (HRP), an enzyme that can catalyze a colorimetric or chemiluminescent reaction, allowing for the visualization and quantification of the target proteins.
Automatically generated - may contain errors
2 protocols using hrp conjugated goat anti rabbit or goat anti mouse secondary antibodies
1
Granulosa Cell Protein Extraction and Western Blot
2
Granulosa Cell Protein Extraction and Western Blot
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The protein was extracted from granulosa cells using RIPA lysis buffer (Beyotime, China) with proteinase and phosphatase inhibitors. The protein concentration was measured by a BCA protein assay kit (Beyotime, China). The samples were denatured at 95 °C for 10 min with 5× protein-loading buffer. The protein was separated by 10% SDS-PAGE and then was transferred onto 0.22-μm PVDF membranes (Millipore, USA). Next, the membranes were blocked with 5% skim milk for 1 h and were then incubated with primary antibodies at 4 °C overnight. Subsequently, the membranes were washed three times and incubated with HRP-conjugated goat anti-rabbit or goat anti-mouse secondary antibodies (1:3000, Abcam, USA) for 1 h. Finally, the proteins were detected by enhanced chemiluminescence and were imaged by an Image Quant LAS 4000 mini biomolecular imager (Bio-Rad, USA).
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