Mouse/Rat Prolactin Antibody

Detection of Mouse and Rat Prolactin by Western Blot. Western blot shows lysates of mouse pituitary tissue and rat pituitary tissue. PVDF membrane was probed with 0.25 µg/mL of Goat Anti-Mouse/Rat Prolactin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1445) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). A specific band was detected for Prolactin at approximately 23 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Cell Proliferation Induced by Prolactin and Neutralization by Mouse Prolactin Antibody. Recombinant Mouse Prolactin (1445-PL) stimulates proliferation in the Nb2-11 rat lymphoma cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Mouse Prolactin (10 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Mouse/Rat Prolactin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1445). The ND₅₀ is typically 0.25-1.0 µg/mL.

Detection of Prolactin in Mouse Pituitary. Prolactin was detected in immersion fixed paraffin-embedded sections of Mouse Pituitary using Goat Anti-Mouse/Rat Prolactin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1445) at 1 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (brown; Catalog # VC004) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in lactotroph cells. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of Mouse and Rat Prolactin by Simple Western^TM. Simple Western shows lysates of mouse pituitary tissue and rat pituitary tissue, loaded at 0.2 mg/ml. A specific band was detected for Prolactin at approximately 26 kDa (as indicated) using 2.5 µg/mL of Goat Anti-Mouse/Rat Prolactin Antigen Affinity-purified Polyclonal Antibody (Catalog # af1445). This experiment was conducted under reducing conditions and using the 12-230kDa separation system.

Detection of Prolactin by Western Blot Overexpression of ER alpha rescues miR-130a-3p-inhibited expression of PRL. The GH3 cells were transfected with the miR-130a-3p mimic and the ER alpha overexpressing vector. The rescue efficiency of ER alpha in the GH3 cells was confirmed by western blotting. (A) The protein expression level of ER alpha was detected by western blotting. GAPDH was used as loading control. (B) The protein expression level of PRL was detected by western blotting. GAPDH was used as loading control. (C) Quantitation of ER alpha protein level. Data are presented as mean ± S.E.M of n = 4 samples per group (P < 0.05 by ANOVA). (D) Quantitation of PRL protein level. Data are presented as mean ± S.E.M of n = 4 samples per group (P < 0.05 by ANOVA). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/32194503), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Prolactin by Western Blot MiR-130a-3p overexpression reduces PRL expression in GH3 cells. GH3 cells were transfected with miR-130a-3p mimic and NC and then the expressions of miR-130a-3p and PRL were analyzed. (A) The expression level of miR-130a-3p was detected by quantitative real-time PCR (qRT-PCR). U6 snRNA was used to normalize the miRNA expression. Data are presented as mean ± S.E.M of n = 6 samples per group (*P < 0.05 by t-test). (B) The expression level of PRL mRNA was detected by qRT-PCR. GAPDH was used to normalize each gene expression. Data are presented as mean ± S.E.M of n = 6 samples per group (*P < 0.05 by t-test). (C) The protein level of PRL in GH3 cells was analyzed by western blotting. GAPDH was used as loading control. (D) Quantitation of the PRL protein level. Data are presented as mean ± S.E.M of n = 4 samples per group (*P < 0.05 by t-test). (E) The protein level of PRL was detected by ICC after GH3 cells transfected with miR-130a-3p mimic and NC. Scale bar, 50 μM. **P < 0.01. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/32194503), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Prolactin by Western Blot Heat stress increases miR-130a-3p expression, reduces PRL and ER alpha expressions in GH3 cells. GH3 cells were separately cultured in 37 or 41°C for 24 h, after that, miR-130a-3p, PRL, and ER alpha expressions were analyzed. (A) The expression level of miR-130a-3p was detected by quantitative real-time PCR (qRT-PCR). U6 snRNA was used to normalize the miRNA expression. Data are presented as mean ± S.E.M of n = 4 samples per group (*P < 0.05 by t-test). (B) The expression levels of HSP70, ER alpha, and PRL mRNAs in GH3 cells were detected by qRT-PCR. GAPDH was used to normalize each gene expression. Data are presented as mean ± S.E.M of n = 4 samples per group (*P < 0.05, ns, not significant by t-test). (C) The protein levels of HSP70, ER alpha, and PRL in GH3 cells were analyzed by western blotting. GAPDH was used as loading control. (D) Quantitation of HSP70, ER alpha, and PRL protein levels. Data are presented as mean ± S.E.M of n = 4 samples per group (*P < 0.05 by t-test). **P < 0.01. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/32194503), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Prolactin by Western Blot Heat stress reduces PRL and ER alpha expressions as well as increases miR-130a-3p expression in the pituitary gland. The mice of treated groups were placed in 40°C for 2 h each time, and the stimulus, respectively lasted 1 and 7 days. The mice in control group were fed as normal in 25°C. Then the expressions of PRL and ER alpha in the pituitary gland were analyzed. (A–B) The mRNA levels of PRL (A) and ER alpha (B) in the pituitary gland of three groups were analyzed by quantitative real-time PCR (qRT-PCR). GAPDH was used to normalize each gene expression. Data are presented as mean ± S.E.M of n = 5 animals per group. Bars that share different letter are significantly different (P < 0.05 by ANOVA). (C) The serum PRL concentration was detected by Elisa assay. Data are presented as mean ± S.E.M of n = 5 animals per group. Bars that share different letter are significantly different (P < 0.05 by ANOVA). (D) The protein levels of PRL and ER alpha in the pituitary gland were analyzed by western blotting. beta -actin was used as loading control. (E) Quantitation of PRL and ER alpha protein levels. Data are presented as mean ± SD of n = 3 animals per group. Bars that share different letter are significantly different (P < 0.05 by ANOVA). The expression levels of miR-130a-3p (F), miR-130b-3p (G), miR-301a-3p (H), and miR-301b-3p (I) were detected by qRT-PCR. U6 snRNA was used to normalize each miRNA expression. Data are presented as mean ± S.E.M of n = 5 animals per group. Bars that share different letter are significantly different (P < 0.05 by ANOVA). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/32194503), licensed under a CC-BY license. Not internally tested by R&D Systems.