|Product Name:||Recombinant Human Oct4_V16-11R Protein|
|Manufacture:||LD Biopharma, Inc.|
Human Oct4 protein encoded by POU5F1 gene is a homeodomain transcription factor of the POU family. This protein is critically involved in the self-renewal of undifferentiated embryonic stem cells. As such, it is frequently used as a marker for undifferentiated cells. Oct4 combined with Sox2, Klf4 and cMyc has been used for generating iPS cells. To further increase Oct4 activity for gene transcription activation, HSV VP16 TAD (transcription activation domain, 76aa) was fused with human Oct4 protein with unique linker domain: L5m.
Recombinant human Oct4_VP16 TAD gene was fully synthesized with codon optimization for E. coli expression. It was constructed with C-terminal tag of 11 arginine domain, which efficiently delivery protein intracellularly. This protein was expressed in E. coli as inclusion bodies, refolded using our unique “temperature shift inclusion body refolding” technology and chromatographically purified. Incubating this protein in various culture mediums at concentration of 2-8 µg/ml may be used for protein derived iPS generation when combined with Sox2, Klf4 and cMyc.
|Accession Number:||NP_002692.2 and NP_044650.1|
|Package Size:||50 µg / Vial|
|Composition:||1.0 mg/ml, sterile-filtered, in 20 mM pH 8.0 Tris-HCl Buffer, with proprietary formulation of NaCl, KCl, EDTA, arginine, DTT and Glycerol.|
|Storage:||In Liquid. Keep at -20°C for long term storage. Product is stable at 4 °C for at least 7 days.|
|Key Reference:||Yang Wang., et al. Reprogramming of mouse and human somatic cells by high-performance engineered factors. EMBO report. vol: 12, pp: 373-378 (2011) Kaija Alfthan1, et al. Properties of a single chain antibody containing different linker peptides. Protein Engineering. Vol: 8. No 7. Pp: 725-731 (1995) Hongyan Zhou, et al. Generation of induced pluripotent stem cells using recombinant protein. Cell Stem Cell. Vol 4. Issue 5: 381-384 (2009)|
1. Protein transduction for protein derived iPS generation.
2. Active recombinant protein, may be used for ELISA based DNA/Protein binding assay.
3. As specific protein substrate for kinase assay.
1. Purity: > 90% by SDS-PAGE.
2. DNA binding assay: DNA binding activity was demonstrated using Oct4 specific DNA binding oligo derived EMSA.
|Recombinant Protein Sequence:||MAGHLASDFAFSPPPGGGGDGPGGPEPGWVDPRTWLSFQGPPGGPGIGPGVGPGSEVWGIPPCPPPYEFCGGMAYCGPQVGVGLVPQGGLETSQPEGEAGVGVESNSDGASPEPCTVTPGAVKLEKEKLEQNPEESQDIKALQKELEQFAKLLKQKRITLGYTQADVGLTLGVLFGKVFSQTTICRFEALQLSFKNMCKLRPLLQKWVEEADNNENLQEICKAETLVQARKRKRTSIENRVRGNLENLFLQCPKPTLQQISHIAQQLGLEKDVVRVWFCNRRQKGKRSSSDYAQREDFEAAGSPFSGGPVSFLLAPGPHFGTPGYGSPHFTALYSSVPFPEGEAFPPVSVTTLGSPMHSNTSRSEPSGPISTGNPSPPSKESHKSPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGGLEESGGGGSPGRRRRRRRRRRR