iMatrix-511 SILK is recombinant human Laminin 511-E8 fragments expressed by transgenic silkworm cocoon. Laminin 511 is known to bind to Intergin α6β1 of the cell surface.
iMatrix-511 SILK can be used as cell culture matrix for various cell types including ES/iPS cells.
Product Size: 1050 ug (175 ug x 6 pcs.)
Format: Liquid Solution (175 ug/vial @0.5 mg/mL), Solvent: PBS (-)
Storage temperature: 2-15 °C
The dissociation constant of the binding activity with integrin α6β1 is under 10nM.
1. Dilute the solution with sterile PBS (-). Coat dishes with 0.5 ug/cm². Add 2 mL of diluted iMatrix-511 solution to the well. The optimal coating concentration depends on cell lines from 0.1 to 1.5 ug/cm². (For example, when you use 6-well plate (9.6 cm²/well), add 9.6 ul of iMatrix-511 (4.8 ug) in 1.99 ml of PBS(-).)
2. Incubate for 1 hour at 37 °C, 3 hours at room temperature, or over night at 4 °C.
3. Remove remaining fluid from the coated surface. No rinse is needed.
4. Immediately plate the cells at desired density.
Notes: Don't allow the plate to dry. Briefly spin down all liquid in the tube before use. Avoid repeated freeze-thaw cycles.
1. Ido, H. et al. The requirement of the glutamic acid residue at the third position from the carboxyl termini of the laminin gamma chains in integrin binding by laminins. J Biol Chem 282, 11144–11154 (2007) doi: 10.1074/jbc.M609402200.
2. Taniguchi, Y. et al. The C-terminal region of laminin beta chains modulates the integrin binding affinities of laminins. J Biol Chem 284, 7820–7831 (2009) doi: 10.1074/jbc.M809332200.
3. Miyazaki, T. et al. Laminin E8 fragments support efficient adhesion and expansion of dissociated human pluripotent stem cells. Nature Communications 3, 1236 (2012) doi: 10.1038/ncomms2231.
4. Nakagawa, M. et al. A novel efficient feeder-free culture system for the derivation of human induced pluripotent stem cells. Scientific Reports 4, 3594 (2014) doi: 10.1038/srep03594.
5. Doi, D. et al. Isolation of Human Induced Pluripotent Stem Cell-Derived Dopaminergic Progenitors by Cell Sorting for Successful Transplantation. Stem Cell Reports 2, 337–350 (2014) doi: 10.1016/j.stemcr.2014.01.013.
6. Takashima, Y. et al. Resetting Transcription Factor Control Circuitry toward Ground-State Pluripotency in Human. Cell 158, 1254–1269 (2014) doi: 10.1016/j.cell.2014.08.029.
7. Fukuta, M. et al. Derivation of Mesenchymal Stromal Cells from Pluripotent Stem Cells through a Neural Crest Lineage using Small Molecule Compounds with Defined Media. PLOS ONE 9, e112291 (2014) doi: 10.1371/journal.pone.0112291.
8. Burridge, P. W. et al. Chemically defined generation of human cardiomyocytes. Nature Methods 11, 855–860 (2014) doi: 10.1038/nmeth.2999.
9. Okumura, N. et al. Laminin-511 and -521 Enable Efficient In Vitro Expansion of Human Corneal Endothelial Cells. Investigative Ophthalmology & Visual Science 56, 2933–2942 (2015) doi: 10.1167/iovs.14-15163.
10. Sasaki, K. et al. Robust In Vitro Induction of Human Germ Cell Fate from Pluripotent Stem Cells. Cell Stem Cell 17, 178–194 (2015) doi: 10.1016/j.stem.2015.06.014.
11. Matsuno, K. et al. Redefining definitive endoderm subtypes by robust induction of human induced pluripotent stem cells. Differentiation 92, 281–290 (2016) doi: 10.1016/j.diff.2016.04.002.
12. Nishimura, K. et al. Estradiol Facilitates Functional Integration of iPSC-Derived Dopaminergic Neurons into Striatal Neuronal Circuits via Activation of Integrin α5β1. Stem Cell Reports 6, 511–524 (2016) doi: 10.1016/j.stemcr.2016.02.008.
13. Takayama, K. et al. Laminin 411 and 511 promote the cholangiocyte differentiation of human induced pluripotent stem cells. Biochemical and Biophysical Research Communications 474, 91–96 (2016) doi: 10.1016/j.bbrc.2016.04.075.
*License to manufacture and sell this product has been obtained from Osaka University & Kyoto University.*
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1 Review Hide Reviews Show Reviews
We've tested both laminin 521 and 511 for various applications, and equivalent adhesion can be seen with the iMatrix fragments at lower concentrations than native laminins. So-far equivalent function at a lower list price per milligram, and lower coating concentrations for applications, for overall substantial cost savings.