After the dewaxing process, muskrat ovaries were slowly washed 3 times with phosphate buffer (0.01M) for 5 minutes each. Samples were added to citrate buffer (10 mM) and heated to repair the antigen, naturally cooled to room temperature then washed as in the previous step. Block with 10% normal goat serum prior to incubation with primary antibody. Incubate the primary polyclonal antibodies against GRP78 (ab21685, Abcam, Cambridgeshire, United Kingdom), P450scc (bs-10099R), 3β-HSD (bs-3906R), P450c17 (bs-3853R), and P450arom (bs-0114R) (Bioss Biotechnology, Beijing, China) at 4°C for 12 hours. After washing off the primary antibody, the tissues were successively incubated with biotin-labeled secondary antibody and horseradish peroxidase working solution (SP-0022, Bioss Biotechnology, Beijing, China) for 30 minutes, respectively, and then visualized with 3, 3-diaminobenzidine (Wako, Tokyo, Japan) solution at room temperature and terminated with distilled water. Finally, the nuclei were re-stained with hematoxylin and tissues were dehydrated and sealed with neutral balsam.
3β hsd bs 3906r
Manufactured by Bioss Antibodies
Sourced in United Kingdom
3β-HSD (bs-3906R) is a laboratory equipment product that functions as an enzyme. It is used in various research applications involving the study of steroidogenesis.
Automatically generated - may contain errors
2 protocols using 3β hsd bs 3906r
1
Immunohistochemical Analysis of Muskrat Ovaries
2
Protein Analysis of Mouse Follicles
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The 100 follicles were pooled from 5 mice at a time for each group. Total protein was extracted from PM and ES follicles using RIPA lysate (P0013B, Beyotime Biotech, Beijing, China) for Western blotting analysis. After SDS‐PAGE, the proteins were transferred to PVDF membrane, which was blocked with 5% skim milk for 1 hour at room temperature. Then, the membrane was incubated with antibodies to FSHR (bs‐20658R; Bioss), Ki‐67 (A2094; ABclonal), 3β‐hydroxysterol dehydrogenase (3β‐HSD) (bs‐3906R; Bioss), P450 17alpha‐hydroxylase (CYP17) (bs‐3853R; Bioss), P450 aromatase (CYP19) (bs‐0114R; Bioss), GAPDH (bs‐2188R; Bioss) or β‐actin (Ac026, ABclonal) overnight at 4°C. After washing, secondary antibody of HRP‐labeled goat anti‐rabbit antibody (bs‐0295G‐HRP; Bioss) was added and incubated for 1 hour at room temperature. Finally, the membrane was subjected to color development and photographed using a Tanon 4600 imaging system. Image was further analyzed by grayscale analysis using Tanon GIS ID software. The experiment for each group was repeated 5 times, and 5 mice were used each time.
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