Product Description

Vitronectin is a 478 amino acid protein (1-19aa = 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 publications indicated that coated recombinant human vitronectin protein alone benefits iPS cell generation when combined with E8 culture medium. The product has also been shown to be an excellent coating matrix material for 1 1R-tagged recombinant TF intracellular delivery for protein derived iPS protocol with extremely low levels of non-specific interaction. 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 a unique “temperature shift inclusion body refolding” technology and chromatographically purified.

Vitronectin is ideal for coating of surfaces. The optimal concentration for cell attachment and culture may differ for various cell types. Vitronectin has been used at a final coating concentration as low as 50 ng/cm2 on plasticware. Further, coating this recombinant protein at 5 – 10 µg/well (6 well plate in either NutriStem or E8 medium can be used for human iPS cell generation in vitro. It is provided in user-friendly packaging for use and storage. Vitronectin is sterile filtered and is supplied as a ready to use solution after thawing and concentration adjustment.

Parameter, Testing, and Method Recombinant Vitronectin #5121
Quantity 0.5 mg
Volume 0.5 mL
Concentration 1 mg/mL
Purity - SDS PAGE Electrophoresis >90%
Formulation 20 mM pH 8 Tris-HCL buffer with proprietary formulation of NaCl, KCl, EDTA, Arginine, DTT and Glycerol
Form Solution
Source Recombinant - E. Coli
Storage Temperature -20°C or -70°C for long term storage
Shelf Life Minimum of 6 months from date of receipt
Sterilization Method Filtration
Cell Attachment Assay Passes
Accession Number NP_000692

Directions for Use

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Coating Procedure:

Use these recommendations as guidelines to determine the optimal coating conditions for your culture system.

  1. Thaw Vitronectin and dilute to desired concentration using serum-free medium or PBS.  The final solution should be sufficiently dilute so that the volume added covers the surface evenly.
  2. Add appropriate amount of diluted material to culture surface.
  3. Incubate at room temperature for 1-2 hours.
  4. Aspirate remaining material.
  5. Rinse plates carefully with dH2O– avoid scratching bottom surface of plates.
  6. Plates are ready for use.  They may also be stored at 2-8°C damp or air dried if sterility is maintained.

Product Applications

Coating recombinant human VTN protein for ES or iPS cell culture can be easily performed as following:

1. Add 1ml PBS buffer to a single well of 6 well plate, and mix well with 10ug recombinant VTN (20ul solution, 0.5mg / ml of VTN stocking solution).

2. Place culture plate at 4 °C for overnight.

3. Plates are ready for routine ES culture. (Remove coating PBS buffer before cell culture).

 

Product References

Vitronectin References:

Wiley, Luke A., et al. "Using patient-specific induced pluripotent stem cells and wild-type mice to develop a gene augmentation-based strategy to treat CLN3-associated retinal degeneration." Human gene therapy 27.10 (2016): 835-846.

Wiley, Luke A., et al. "Generation of xeno‐free, cGMP‐compliant patient‐specific iPSCs from skin biopsy." Current protocols in stem cell biology 42.1 (2017): 4A-12.

Dwyer, Sheila Figel, Lingqiu Gao, and Irwin H. Gelman. "Identification of novel focal adhesion kinase substrates: Role for FAK in NFκB signaling." International journal of biological sciences 11.4 (2015): 404.

Wiley, Luke A., et al. "cGMP production of patient-specific iPSCs and photoreceptor precursor cells to treat retinal degenerative blindness." Scientific reports 6 (2016): 30742.

Kittur, Harsha, et al. "Probing Cell Adhesion Profiles with a Microscale Adhesive Choice Assay." Biophysical journal 113.8 (2017): 1858-1867.

Dwyer, Sheila Figel, and Irwin H. Gelman. "Cross-phosphorylation and interaction between Src/FAK and MAPKAP5/PRAK in early focal adhesions controls cell motility." Journal of cancer biology & research 2.1 (2014).

Worthington, Kristan S., et al. "Two-photon polymerization for production of human iPSC-derived retinal cell grafts." Acta biomaterialia 55 (2017): 385-395.

Product Certificate of Analysis

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Safety and Documentation

Safety Data Sheet

Certificate of Origin

Product Disclaimer

This product is for R&D use only and is not intended for human or other uses. Please consult the Material Safety Data Sheet for information regarding hazards and safe handling practices.