Generation of monoclonal antibodies to the zinc finger domain of the eukaryotic transcription factor Sp1

Hitoshi Katai, Kuniaki Terato, Rajendra Raghow

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Using a synthetic peptide that encompasses the zinc finger domain of the eukaryotic transcription factor Sp1, we produced a number of monoclonal antibodies that specifically reacted with the target antigen. Analysis by competitive inhibition assay of five of the monoclonal antibodies revealed that they all recognized a dominant epitope in the synthetic peptide and reacted strongly to recombinantly synthesized β-galactosidase-Sp1 fusion polypeptide. To determine cellular distribution of Sp1-like molecules, cytoplasmic and nuclear proteins from human lung fibroblasts (HFL-1) and a human rhabdomyosarcoma cell line (A204) were immunoblotted and reacted with our antibodies. In addition to the well characterized 95 Kd and 105 Kd proteins, considered to be the authentic Sp1 polypeptide, a number of other cellular proteins reacted with these antibodies. Immunofluorescence staining of the cells with mAb to the zinc finger of Sp1 also revealed cell-specific differences in intracellular distribution of Sp1-like molecules. Both cytoplasmic and nuclear staining was readily observed in the rhabdomyosarcoma cells. In contrast, while some HFL-1 cells exhibited staining of only cytoplasm, both cytoplasmic and nuclear immunofluorescence was seen in others.

Original languageEnglish (US)
Pages (from-to)73-81
Number of pages9
JournalMolecular and Cellular Biochemistry
Volume101
Issue number1
DOIs
StatePublished - Feb 1 1991

Fingerprint

Sp1 Transcription Factor
Zinc Fingers
Zinc
Monoclonal Antibodies
Peptides
Rhabdomyosarcoma
Staining and Labeling
Fluorescent Antibody Technique
A 204
Galactosidases
Molecules
Antibodies
Fibroblasts
Nuclear Proteins
Epitopes
Assays
Cytoplasm
Proteins
Fusion reactions
Cells

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Clinical Biochemistry
  • Cell Biology

Cite this

Generation of monoclonal antibodies to the zinc finger domain of the eukaryotic transcription factor Sp1. / Katai, Hitoshi; Terato, Kuniaki; Raghow, Rajendra.

In: Molecular and Cellular Biochemistry, Vol. 101, No. 1, 01.02.1991, p. 73-81.

Research output: Contribution to journalArticle

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