Nacalai USA, Inc. | Product | Vitronectin-398™ (Xeno-free)

Vitronectin-398™ (Xeno-free)

ES Quality Recombinant Human

Human VTN (Vitronectin) is a 478 amino acid protein (1-19 = signal domain) that belongs to a member of the pexin family. Vitronectin is an abundant glycoprotein found in serum and the extracellular matrix. It promotes cell adhesion and spreading, inhibits the membrane-damaging effect of the terminal cytolytic complement pathway, and binds to several serpin serine protease inhibitors. It is a secreted protein and exists in either a single chain form or a clipped, two chain form held together by a disulfide bond. Vitronectin has been speculated to be involved in hemostasis and tumor malignancy. Recent publication from James Thomson's group indicated that coated recombinant human vitronectin protein alone benefits iPS cell generation when combined with E8 culture medium.

Features

✔ Non-Fusion Protein
✔ User Friendly Protocol
✔ Cost Effective

Human ES cells (H1) were cultivated in xeno-free medium (NutriStem™) on Vitronectin-398™ (Xeno-Free) coated 6-well plate for 10 generations, and staining With Oct4, TRA-81 and DAPI.

Domain Structure

Recombinant human Vitronectin gene (20-398 aa Fragment) was constructed with codon optimization and expressed in non-fusion protein form in E.coli as inclusion bodies. The final product was refolded using our unique "temperature shift inclusion body refolding" technology and chromatographically purified. Coating this recombinant protein at 5-10 µg / well (6 well plate) in either NutirStem or E8 medium can be used for human iPS cell generation in vitro.

Reference

Guokai Chen, et al. Cehmically defined conditions for human iPSC derivation and culture.
Nature Methods. 8, 424-429 (2011)

Stefan R. Braam. Et al. Recombinant Vitronectin is a Fucntionally Defined Substrate That Supports Human Embryonic Stem Cell Self-Renewal via aVb5 integrin.
STEM CELLS. Vol 26. Issue 9. 2257-2265 (2008)

Application

1. As coating matrix protein for maintaining long-term ES or iPS cell culture before combining with E8 culture medium.

2. As an excellent coating matrix material of 11R-tagged recombinant TF intracellular delivery for protein derived iPS protocol with extremely low-level non-specific interaction.

Coating Protocol

Use 1 ml PBS per well, add 5-10 µg protein to each well and incubate at 4°C overnight.
After coating, remove PBS solution, the plate is ready for ES cell cultivation.

Accession Number

NP_000629

Species

Human

Purity

>95% by SDS-PAGE

Functional Test

Each lot was tested with human ES cell (H1) culture using 5-10 µg in 1 ml Nutristem medium per well (6-well plate).

Composition

0.5 mg/ml, sterile-filtered, in 20 mM pH 8.0 Tris-HCl Buffer, with proprietary formulation buffer containing sucrose, EDTA, Arginine and DTT.

Storage

-80℃ for long term storage.

Recombinant Protein Sequence

MDQESCKGRCTEGFNVDKKCQCDELCSYYQSCCTDYTAECKPQVTRGDVFTMPEDEYTVYDDGE EKNNATVHEQVGGPSLTSDLQAQSKGNPEQTPVLKPEEEAPAPEVGASKPEGIDSRPETLHPGR PQPPAEEELCSGKPFDAFTDLKNGSLFAFRGQYCYELDEKAVRPGYPKLIRDVWGIEGPIDAAF TRINCQGKTYLFKGSQYWRFEDGVLDPDYPRNISDGFDGIPDNVDAALALPAHSYSGRERVYFF KGKQYWEYQFQHQPSQEECEGSSLSAVFEHFAMMQRDSWEDIFELLFWGRTSAGTRQPQFISRD WHGVPGQVDAAMAGRIYISGMAPRPSLAKKQRFRHRNRKGYRSQRGHSRGRNQNSRRPSR

Ordering Information

Product	Cat.No.	Storage	PKG Size	Price(US$)
Vitronectin-398™ (Xeno-free), Recombinant Human	NU0006	-20°C	500 µg	99.00	Buy