Purification and Quality Control Purified protein is greater than 95% homogeneous based on SDS-PAGE analysis.
Unit Definition (Activity) 1 unit equals 1 nanogram of protein. 2-10 units are sufficient for a gel mobility shift assay in a 20µl reaction. 50-200 units are required for a reconstituted transcription assay and 100 units are sufficient for a protein-protein interaction assay. variable in different lots
Applications The N-terminal Flag-tagged WT-1 protein (residue 1-446, including exon 5 but without KTS) was expressed in a baculovirus system and purified by affinity and FPLC chromatography. Recombinant WT-1 protein can be used for: 1) in vitro function studies including transcription, DNA or RNA binding assays; 2) protein-protein interaction assay; and 3) cell growth and proliferation assays.
Formulation and Storage 1x dilution buffer A: 20 mM Tris-Cl (pH 8.0), 20% Glycerol, 100 mM KCl, 1 mM DTT and 0.2 mM EDTA
Synonym
Protein Sequence
Background WT-1, the product of Wilms’ tumor suppressor gene Wt1, is a nuclear protein with structural motifs characteristic of transcription factors, including four C-terminal zinc fingers. While different pre-mRNA processing could result in 16 isoforms of the protein, inclusion or exclusion of exon 5 and the three amino acids (KTS) between zinc fingers 3 and 4 largely affects the activity of WT1 protein (1-3). Such a complex post-transcriptional regulation, particularly in splicing, may represent a major regulatory mechanism for tumorigenesis of the Wilms’ tumor. WT1 (-KTS) appears to have different binding affinity to both DNA and RNA comparing to the +KTS form (4, 5).
References: 1. Haber, DA. et al., (1991) Proc. Natl. Acad. Sci. USA 88, 9618-9622; (1993) Science 262, 2057-2059 2. Kreidberg, JA. et al., (1993) Cell 74, 679-691 3. Larsson, SH. et al., (1995) Cell 81, 391-401 4. Little, M. et al., (1997) Hum. Mutat. 9, 209-225 5. Englert, C. et al., (1995) EMBO J. 14, 4662-4675
DISCLAIMER
This products is recommended For RESEARCH USE ONLY and is Not qualified for Use in Diagnostic or Therapeutic Procedures.
