Avian stem cell factor (SCF): production and characterization of the recombinant His-tagged SCF of chicken and its neutralizing antibody

Avian stem cell factor (SCF): production and characterization of the recombinant His-tagged SCF of chicken and its neutralizing antibody

Recombinant chicken stem cell factor (SCF) was produced in bacteria as a histidine-tagged protein (HisΔSCF) and purified by affinity chromatography on an Ni 2+-NTA agarose column. HisΔSCF is devoid of signal peptide and transmembrane domains and therefore mimics a soluble form of SCF. We demonstrate...

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Bibliographic Details
Published in:Cytokine (Philadelphia, Pa.) Vol. 8; no. 1; pp. 14 - 20
Main Authors: BartňÏk, Petr, Pichlı́ková, Lenka, Stengl, Gabriele, Boehmelt, Guido, Martin, Frank H., Beug, Hartmut, Dvōák, Michal, Zenke, Martin
Format: Journal Article
Language:English
Published: England Elsevier Ltd 1996
Subjects:
Amino Acid Sequence
Animals
Antibodies
Cell Differentiation
Cell Division - drug effects
Cell Line
chicken
Chickens
DNA Primers
E. coliexpression
Erythrocytes - cytology
Hematopoietic Stem Cells - cytology
Hematopoietic Stem Cells - drug effects
Histidine
histidine tag
Molecular Sequence Data
Neutralization Tests
Polymerase Chain Reaction
Proto-Oncogene Proteins c-kit - physiology
Rabbits
Recombinant Proteins - biosynthesis
Recombinant Proteins - immunology
Recombinant Proteins - pharmacology
SCF
Sequence Tagged Sites
stem cell factor
Stem Cell Factor - biosynthesis
Stem Cell Factor - immunology
Stem Cell Factor - pharmacology
Thymidine - metabolism
Transfection
histidine tag
chicken
stem cell factor
E. coliexpression
SCF
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Summary:Recombinant chicken stem cell factor (SCF) was produced in bacteria as a histidine-tagged protein (HisΔSCF) and purified by affinity chromatography on an Ni 2+-NTA agarose column. HisΔSCF is devoid of signal peptide and transmembrane domains and therefore mimics a soluble form of SCF. We demonstrate that the factor is biologically fully active in supporting sustained proliferation of SCF-dependent red blood progenitor cells of chicken in vitro. Polyclonal SCF-specific antibodies were raised in rabbits and shown to display potent SCF-neutralizing activity. Thus, recombinant chicken HisΔSCF together with the SCF-specific antibody provide valuable tools for studying the role of SCF and c-kit receptor in development, growth and differentiation of avian haematopoietic cells.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
ISSN:1043-4666
1096-0023
DOI:10.1006/cyto.1996.0003