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

Melwyn Sequeira, Farakid Al-Khafaji, SooMi Park, Mark D. Lewis, Malcolm H. Wheeler, V. Krishna K Chatterjee, Bharat Jasani, Marian Ludgate

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)927-932
Number of pages6
JournalThyroid
Volume13
Issue number10
Publication statusPublished - Oct 2003
Externally publishedYes

Fingerprint

Hair Follicle
Testis
Thyroid Gland
Transcription Factors
Antibodies
Proteins
Thyroid Dysgenesis
Staining and Labeling
human FOXE1 protein
Seminiferous Tubules
Germ-Line Mutation
Cleft Palate
Nickel
Cell Nucleus
Paraffin
Connective Tissue
Hair
Formaldehyde
Endothelium
Smooth Muscle

ASJC Scopus subject areas

  • Endocrinology

Cite this

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. / Sequeira, Melwyn; Al-Khafaji, Farakid; Park, SooMi; Lewis, Mark D.; Wheeler, Malcolm H.; Chatterjee, V. Krishna K; Jasani, Bharat; Ludgate, Marian.

In: Thyroid, Vol. 13, No. 10, 10.2003, p. 927-932.

Research output: Contribution to journalArticle

Sequeira, M, Al-Khafaji, F, Park, S, Lewis, MD, Wheeler, MH, Chatterjee, VKK, Jasani, B & Ludgate, M 2003, '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', Thyroid, vol. 13, no. 10, pp. 927-932.
Sequeira, Melwyn ; Al-Khafaji, Farakid ; Park, SooMi ; Lewis, Mark D. ; Wheeler, Malcolm H. ; Chatterjee, V. Krishna K ; Jasani, Bharat ; Ludgate, Marian. / 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. In: Thyroid. 2003 ; Vol. 13, No. 10. pp. 927-932.
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