Production and Application of Polyclonal Antibody to Human Thyroid Transcription Factor 2 Reveals Thyroid Transcription Factor 2 Protein Expression in Adult Thyroid and Hair Follicles and Prepubertal Testis
摘要:
Germline mutations in thyroid transcription factor 2 (TTF2) cause thyroid agenesis, spiky hair, and cleft palate, indicating thyroidal and extrathyroidal expression. We sought to investigate this by producing and applying an antibody to human TTF2. The coding region of human TTF2 was cloned into a bacterial expression vector, production of the soluble TTF2 protein optimized, and pure TTF2 obtained by nickel chromatography. Rabbits were immunized and the resulting TTF2 polyclonal titrated on formalin-fixed, paraffin-embedded sections of thyroid. The optimized protocol was applied to a range of tissues. Nine milligrams of TTF2 protein was obtained per liter of culture and a high-titer antibody produced. This displayed specific staining of thyroid follicular cell nuclei/cytoplasm and not of the interstitium, connective tissue, smooth muscle, or endothelium. No staining was obtained with the preimmune serum in the same conditions, or with the majority of other tissues tested with the TTF2 polyclonal. The exceptions were testis and skin, in which nuclear TTF2 immunoreactivity was present in the seminiferous tubules and cells in the follicular outer root sheath, respectively. In conclusion, we have produced a polyclonal antibody for human TTF2 and demonstrated immunoreactivity for this transcription factor in adult human thyroid and hair follicles and prepubertal testis.
Fluorosteroids VII. Synthesis and biological activity of derivatives of 6α-fluoro-16α,17α-isopropylidenedihydroxypregna-1,4-diene-11β, 21-diol--3,20-dione
Cesium Fluoride Catalyzed Trifluoromethylation of Esters, Aldehydes, and Ketones with (Trifluoromethyl)trimethylsilane
作者:Rajendra P. Singh、Ganfeng Cao、Robert L. Kirchmeier、Jean'ne M. Shreeve
DOI:10.1021/jo982494c
日期:1999.4.1
low reactivity of carboxylic esters toward (trifluoromethyl)trimethylsilane (TMS-CF(3)) was investigated. A universal cesiumfluoridecatalyzed procedure for nucleophilic trifluoromethylation was developed. At room temperature (25 degrees C), with catalytic amounts of cesiumfluoride, carboxylic esters were found to react to give the silyl ether intermediates, which afforded the trifluoromethyl ketones