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Fig. 3 | Journal of Ovarian Research

Fig. 3

From: Case report: Birth achieved after effective ovarian stimulation combined with dexamethasone in a patient with resistant ovary syndrome

Fig. 3

Immunoprecipitation followed by western blotting and dot blot analysis of anti-FSHR autoantibodies in the serum of the patient. A polyclonal antibody against FSHR (Abcam, ab113421) in TBST buffer (containing 3% BSA) was used as positive control serum with anti-FSHR autoantibodies, a healthy person’s serum was used as negative control, and sera from the patient before and after the dexamethasone treatment were used to immunoprecipitate recombinant FSHR protein purified from E. coli (ImmunoClone, IC8974-A). Samples comprising 0.6 μg of FSHR protein per well were separated on 10% SDS-PAGE gels and then transferred to nitrocellulose membranes. For the dot blot assay, FSHR protein (100 ng/dot) was applied onto nitrocellulose membranes. The membranes were incubated overnight at 4 °C with serum from the patient and the healthy control, followed by incubation with a horseradish peroxidase (HRP)-conjugated anti-human secondary antibody (Proteintech, SA00001–11) or anti-rabbit secondary antibody (Proteintech, SA00001–2). Stained bands or dots were visualized using Omni-ECL reagent (EpiZyme, SQ201). PC: positive control; BD: before the dexamethasone treatment; AD: after the dexamethasone treatment. NC: negative control; FSHR: follicle-stimulating hormone receptor

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