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 (His delta SCF) and purified by affinity chromatography on an Ni(2+)-NTA agarose column. His delta SCF 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 His delta SCF 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.
Bartůnĕk
P
P
Institute of Molecular Pathology, Vienna, Austria.
Pichlíková
L
L
Stengl
G
G
Boehmelt
G
G
Martin
F H
FH
Beug
H
H
Dvorák
M
M
Zenke
M
M
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D002454:Cell Differentiation; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002645:Chickens; D017931:DNA Primers; D004912:Erythrocytes / Q000166:cytology; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000187:drug effects; D006639:Histidine; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D016133:Polymerase Chain Reaction; D019009:Proto-Oncogene Proteins c-kit / Q000502:physiology; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000276:immunology / Q000494:pharmacology; D016324:Sequence Tagged Sites; D019089:Stem Cell Factor / Q000096:biosynthesis* / Q000276:immunology / Q000494:pharmacology; D013936:Thymidine / Q000378:metabolism; D014162:Transfection
D000906:Antibodies; D017931:DNA Primers; D011994:Recombinant Proteins; D019089:Stem Cell Factor; D006639:Histidine; D019009:Proto-Oncogene Proteins c-kit; D013936:Thymidine