Production of a specific polyclonal antibody against the rat β thyroid receptor, using synthetic peptide as antigen

Abstract

Abstract. We have developed a polyclonal antibody to a synthetic peptide corresponding to the deduced amino acid residues 67–80 of the amino terminal sequence of the rat thyroid receptor β cDNA. Immunopositive sera were identifed by radioimmunoassay and purified by affinity chromatography. The antibody specifically immunoprecipitated the labelled thyroid receptor β synthetized in vitro but not the rat thyroid receptor α. Furthermore, 38% of the L-T3 binding capacity of the rat liver nuclear receptor were immunoabsorbed by this antibody. By immunocytochemistry, this antibody stained specifically the nuclei of hepatocytes while no staining was observed in the rat testis. This study presents a specific polyclonal antibody against the thyroid receptor β. This antibody will be of a great help in determining the role of the β receptors in the mechanism of action of thyroid hormone.

Coexpression of thyroid hormone receptor isoforms in mouse oligodendrocytes

“…One peptide corresponding to residues 71–85 of the rat β1‐TR N terminal region and another to residues 12–23 of the N terminal region common to rat α1‐ and α2‐TR (α‐TR) were synthesized (IGBMC, Strasbourg‐Illkirch, France), checked by high‐performance liquid chromatography (HPLC) for purity (>95%), and coupled to keyhole limpet hemocyanin (KLH; Megathura crenulata ; Sigma‐Aldrich Chimie France), using m‐maleimidobenzoic acid N‐hydroxy‐succinimide ester as previously described (Puymirat et al, 1991).…”

Thyroid hormone up‐regulates thyroid hormone receptor β gene expression in rat cerebral hemisphere astrocyte cultures

“…The preparation and characterization of the p l thyroid receptor antibodies has been described in previous work (Puymirat et al, 1991(Puymirat et al, ,1992a. Cultures were fixed with 1% paraformaldehyde in 0.1 M sodium phosphate buffer, 0.2% NaCl, pH 7.4 (PBS) for 15 min at room temperature.…”

“…Cultures were fixed with 1% paraformaldehyde in 0.1 M sodium phosphate buffer, 0.2% NaCl, pH 7.4 (PBS) for 15 min at room temperature. Cultures were then washed with PBS, permeabilized for 10 min with 0.2% Triton X-100, and incubated with affinity-purified anti-pl antibodies (10 pg/ml) for 2 h at room temperature (Puymirat et al, 1991). After washing with PBS, cells were incubated with an FITC goat anti-rabbit IgG (1150, obtained from Boehringer).…”

Targeting nuclear receptors for NASH/MASH: From bench to bedside

“…13 In the liver, THRs are expressed in hepatocytes, cholangiocytes, kupffer cells, and HSCs, and therefore, TH can influence almost all aspects of hepatic physiology. 14 – 17 Epidemiological studies have shown an increased incidence of MASLD with both overt and subclinical hypothyroidism in humans. 18 – 23 Furthermore, animal models of MASH have shown evidence of intra-hepatic TH insufficiency, suggesting deregulated TH metabolism in fatty liver.…”