Journal: Journal of the Endocrine Society

Article Title: Validation of a new Custom Polyclonal Progesterone Receptor Antibody for Immunohistochemistry in the Female Mouse Brain

doi: 10.1210/jendso/bvad113

Figure Lengend Snippet: Immunoreactivity produced by commercial and custom PR antibodies in the arcuate nucleus. (A) Schematic diagram of the human PR-A and PR-B protein amino acid sequences, with the corresponding immunogen sequences of tested PR antibodies mapped. Host species of antibodies is indicated by r (rabbit) or m (mouse). AF, activation functions; DBD, DNA-binding domain; h, hinge region; LBD, ligand-binding domain; NTD, N-terminal domain. (B-H) Representative images of chromogenic immunohistochemical labeling in the arcuate nucleus of adult diestrous female mice produced by the (B) discontinued A0098 DAKO PR antibody, (C-G) the commercially available MA1-410 (1:5000), PA1413 (1:500), bs-0111R (1:1000), and MAB9785 (1:2000) PR antibodies, and (H) the newly generated custom RC269 PR antibody (1:2500) on paraformaldehyde-fixed brain sections. (F, G) Representative images of the high-signal (n = 3 mice) and low-signal (n = 3) MAB9785 PR antibody labeling observed across the 6 animals tested. Scale bar = 100 µm. 3 V, third ventricle; ARN, arcuate nucleus.

Article Snippet: PA1413 Boster Bio (Pleasanton, CA, USA) , Rabbit polyclonal , 536-553aa N-terminal domain (close to the DNA-binding domain) , AB_2923359 , Unclear , [ ] .

Techniques: Produced, Activation Assay, Binding Assay, Ligand Binding Assay, Immunohistochemical staining, Labeling, Generated, Antibody Labeling

Journal: Animal Reproduction

Article Title: Regulation of progesterone during follicular development by FSH and LH in sheep

doi: 10.1590/1984-3143-AR2022-0027

Figure Lengend Snippet: Primers used for qRT‐PCR.

Article Snippet: The electrotransfer solution was washed away with 1× trimethylaminomethane buffer saline (TBST) and was sealed with TBST which contained 5% skimmed milk powder for 1 h. The antibody was incubated with rabbit polyclonal antibody StAR (bs-3570R, 1:500, Bioss) P450scc (bs-10099R, 1:500, Bioss) PGR (bs-23376R, 1:500, Bioss) with β-actin (bs-0061R, 1:3,000, Bioss) as an internal reference, overnight at 4 °C, after washing, goat anti-rabbit secondary antibody (bs-0295G-HRP, Bioss) at a dilution ratio of 1:3,000, at 37 °C for 1 h. Immune complexes were detected with enhanced chemiluminescence solution (Abnova, Taibei, Taiwan), and the expression was quantified using ImageJ (National Institutes of Health).

Techniques:

Journal: Animal Reproduction

Article Title: Regulation of progesterone during follicular development by FSH and LH in sheep

doi: 10.1590/1984-3143-AR2022-0027

Figure Lengend Snippet: (A) shows the results of Hemathoxylin/Eosin (HE) staining and (B) Negative control. Immunolocalization of StAR (C), P450scc (D) and PGR (E) proteins in sheep follicles; (F-H) shows the immunofluorescence localization of PGR (green) in granulosa cells, and the nuclei are restained with DAPI (blue). Original magnification: Cellular immunofluorescence and immunohistochemistry: x400, HE and negative control: x200. OC: oocyte; CCs: cumulus cells; GCs: granulosa cells; TCs: theca cells.

Article Snippet: The electrotransfer solution was washed away with 1× trimethylaminomethane buffer saline (TBST) and was sealed with TBST which contained 5% skimmed milk powder for 1 h. The antibody was incubated with rabbit polyclonal antibody StAR (bs-3570R, 1:500, Bioss) P450scc (bs-10099R, 1:500, Bioss) PGR (bs-23376R, 1:500, Bioss) with β-actin (bs-0061R, 1:3,000, Bioss) as an internal reference, overnight at 4 °C, after washing, goat anti-rabbit secondary antibody (bs-0295G-HRP, Bioss) at a dilution ratio of 1:3,000, at 37 °C for 1 h. Immune complexes were detected with enhanced chemiluminescence solution (Abnova, Taibei, Taiwan), and the expression was quantified using ImageJ (National Institutes of Health).

Techniques: Staining, Negative Control, Immunofluorescence, Immunohistochemistry

Journal: Animal Reproduction

Article Title: Regulation of progesterone during follicular development by FSH and LH in sheep

doi: 10.1590/1984-3143-AR2022-0027

Figure Lengend Snippet: P4 levels, StAR, P450scc, 3β-HSD and PGR levels in different developmental stages of follicles. (A) P4 levels at different follicular development stages; (B) The relative gene levels of StAR, P450scc, 3β-HSD and PGR; (C) Western blot of StAR, P450scc and PGR; (D) the relative protein levels of StAR, P450scc and PGR. β-Actin was used as internal control. Values indicate means ± SEM. * p < 0.05 compared with large follicle group. PGR: Progesterone receptor; SEM: standard error of mean

Article Snippet: The electrotransfer solution was washed away with 1× trimethylaminomethane buffer saline (TBST) and was sealed with TBST which contained 5% skimmed milk powder for 1 h. The antibody was incubated with rabbit polyclonal antibody StAR (bs-3570R, 1:500, Bioss) P450scc (bs-10099R, 1:500, Bioss) PGR (bs-23376R, 1:500, Bioss) with β-actin (bs-0061R, 1:3,000, Bioss) as an internal reference, overnight at 4 °C, after washing, goat anti-rabbit secondary antibody (bs-0295G-HRP, Bioss) at a dilution ratio of 1:3,000, at 37 °C for 1 h. Immune complexes were detected with enhanced chemiluminescence solution (Abnova, Taibei, Taiwan), and the expression was quantified using ImageJ (National Institutes of Health).

Techniques: Western Blot

Journal: Animal Reproduction

Article Title: Regulation of progesterone during follicular development by FSH and LH in sheep

doi: 10.1590/1984-3143-AR2022-0027

Figure Lengend Snippet: Effect of different concentrations of FSH on P4 secretion (A), gene (B) and protein expression (C, D) of StAR, P450scc, 3β-HSD and PGR in cultured GCs. β-Actin was used as internal control. β-Actin was used as internal control. Values indicate means ± SEM. * p < 0.05 compared with control group. FSH: follicle stimulating hormone; SEM: standard error of mean.

Article Snippet: The electrotransfer solution was washed away with 1× trimethylaminomethane buffer saline (TBST) and was sealed with TBST which contained 5% skimmed milk powder for 1 h. The antibody was incubated with rabbit polyclonal antibody StAR (bs-3570R, 1:500, Bioss) P450scc (bs-10099R, 1:500, Bioss) PGR (bs-23376R, 1:500, Bioss) with β-actin (bs-0061R, 1:3,000, Bioss) as an internal reference, overnight at 4 °C, after washing, goat anti-rabbit secondary antibody (bs-0295G-HRP, Bioss) at a dilution ratio of 1:3,000, at 37 °C for 1 h. Immune complexes were detected with enhanced chemiluminescence solution (Abnova, Taibei, Taiwan), and the expression was quantified using ImageJ (National Institutes of Health).

Techniques: Expressing, Cell Culture

Journal: Animal Reproduction

Article Title: Regulation of progesterone during follicular development by FSH and LH in sheep

doi: 10.1590/1984-3143-AR2022-0027

Figure Lengend Snippet: Effect of different concentrations of LH on P4 secretion (A), gene (B) and protein expression (C-D) of StAR, P450scc, 3β-HSD and PGR in cultured GCs. β-Actin was used as internal control. Values indicate means ± SEM. * p < 0.05 compared with control group. LH: luteinizing hormone; SEM: standard error of mean.

Article Snippet: The electrotransfer solution was washed away with 1× trimethylaminomethane buffer saline (TBST) and was sealed with TBST which contained 5% skimmed milk powder for 1 h. The antibody was incubated with rabbit polyclonal antibody StAR (bs-3570R, 1:500, Bioss) P450scc (bs-10099R, 1:500, Bioss) PGR (bs-23376R, 1:500, Bioss) with β-actin (bs-0061R, 1:3,000, Bioss) as an internal reference, overnight at 4 °C, after washing, goat anti-rabbit secondary antibody (bs-0295G-HRP, Bioss) at a dilution ratio of 1:3,000, at 37 °C for 1 h. Immune complexes were detected with enhanced chemiluminescence solution (Abnova, Taibei, Taiwan), and the expression was quantified using ImageJ (National Institutes of Health).

Techniques: Expressing, Cell Culture

Journal: Animal Reproduction

Article Title: Regulation of progesterone during follicular development by FSH and LH in sheep

doi: 10.1590/1984-3143-AR2022-0027

Figure Lengend Snippet: Effects of FSH+LH on P4 secretion and the levels of StAR, P450scc, 3β‐HSD and PGR in cultured GCs. (A) P4 levels; (B) P4 synthetase and receptor relative gene expression; (C, D) Western blotting and relative protein expression of P4 synthetase and receptor. β-Actin was used as internal control. Values indicate means ± SEM. * p < 0.05 compared with control group. F1: FSH (1IU/mL); LH: luteinizing hormone; FSH: follicle stimulating hormone; SEM: standard error of mean.

Article Snippet: The electrotransfer solution was washed away with 1× trimethylaminomethane buffer saline (TBST) and was sealed with TBST which contained 5% skimmed milk powder for 1 h. The antibody was incubated with rabbit polyclonal antibody StAR (bs-3570R, 1:500, Bioss) P450scc (bs-10099R, 1:500, Bioss) PGR (bs-23376R, 1:500, Bioss) with β-actin (bs-0061R, 1:3,000, Bioss) as an internal reference, overnight at 4 °C, after washing, goat anti-rabbit secondary antibody (bs-0295G-HRP, Bioss) at a dilution ratio of 1:3,000, at 37 °C for 1 h. Immune complexes were detected with enhanced chemiluminescence solution (Abnova, Taibei, Taiwan), and the expression was quantified using ImageJ (National Institutes of Health).

Techniques: Cell Culture, Expressing, Western Blot

Journal: Animal Reproduction

Article Title: Regulation of progesterone during follicular development by FSH and LH in sheep

doi: 10.1590/1984-3143-AR2022-0027

Figure Lengend Snippet: Effects of FSH+LH on P4 secretion and the levels of StAR, P450scc, β‐HSD and PGR in cultured GCs. (A) P4 levels; (B) P4 synthetase and receptor relative gene expression; (C, D) Western blotting and relative protein expression of P4 synthetase and receptor. Values indicate means ± SEM. * p < 0.05 compared with control group. F0.1: FSH (0.1IU/mL); LH: luteinizing hormone; FSH: follicle stimulating hormone; SEM: standard error of mean

Article Snippet: The electrotransfer solution was washed away with 1× trimethylaminomethane buffer saline (TBST) and was sealed with TBST which contained 5% skimmed milk powder for 1 h. The antibody was incubated with rabbit polyclonal antibody StAR (bs-3570R, 1:500, Bioss) P450scc (bs-10099R, 1:500, Bioss) PGR (bs-23376R, 1:500, Bioss) with β-actin (bs-0061R, 1:3,000, Bioss) as an internal reference, overnight at 4 °C, after washing, goat anti-rabbit secondary antibody (bs-0295G-HRP, Bioss) at a dilution ratio of 1:3,000, at 37 °C for 1 h. Immune complexes were detected with enhanced chemiluminescence solution (Abnova, Taibei, Taiwan), and the expression was quantified using ImageJ (National Institutes of Health).

Techniques: Cell Culture, Expressing, Western Blot

Journal: Animal Reproduction

Article Title: Regulation of progesterone during follicular development by FSH and LH in sheep

doi: 10.1590/1984-3143-AR2022-0027

Figure Lengend Snippet: Effects of FSH+LH on P4 secretion and the levels of StAR, P450scc, β‐HSD and PGR in cultured GCs. (A) P4 levels; (B) P4 synthetase and receptor relative gene expression; (C, D) Western blotting and relative protein expression of P4 synthetase and receptor. Values indicate means ± SEM. * p < 0.05 compared with control group. F0.01: FSH (0.01IU/mL); LH: luteinizing hormone; FSH: follicle stimulating hormone; SEM: standard error of mean.

Article Snippet: The electrotransfer solution was washed away with 1× trimethylaminomethane buffer saline (TBST) and was sealed with TBST which contained 5% skimmed milk powder for 1 h. The antibody was incubated with rabbit polyclonal antibody StAR (bs-3570R, 1:500, Bioss) P450scc (bs-10099R, 1:500, Bioss) PGR (bs-23376R, 1:500, Bioss) with β-actin (bs-0061R, 1:3,000, Bioss) as an internal reference, overnight at 4 °C, after washing, goat anti-rabbit secondary antibody (bs-0295G-HRP, Bioss) at a dilution ratio of 1:3,000, at 37 °C for 1 h. Immune complexes were detected with enhanced chemiluminescence solution (Abnova, Taibei, Taiwan), and the expression was quantified using ImageJ (National Institutes of Health).

Techniques: Cell Culture, Expressing, Western Blot