Description
General description
Cardiomyocytes are the cardiac muscle cells that make up the heart and are responsible for generating contractile forces within the heart. Laminin-221 is known to bind to the integrin α7X2β1 which is located on the cell surface of pluripotent stem cells and is known to induce cardiac and skeletal muscle differentaition. Professor Kiyotoshi Sekiguchi′s group (Matrixome, Inc.) have produced a recombinant E8 fragment of laminin-221 at large-scale while retaining the full integrin binding activity. ECMatrix-221 E8 Laminin Substrates are recombinant Laminin-221-E8 fragments which bind to integrin α7X2β1 and increases the differentiation of pluripotent stem cells (ES/iPSCs) into functional cardiomyocytes.
Application
Xeno-free laminin-221 coating for the differentiation of pluripotent stem cells into cardiomyocytes, 1050 μg (CHO-S derived)
Preparation Note
ECMatrix-221 E8 Laminin Substrates should be stored at 2-8°C. Avoid multiple freeze-thaw cycles and protect from light.
Precoating Method
1. Dilute the 0.5 mg/mL stock solution with sterile PBS to achieve a 2.5 μg/mL working solution.
2. Coat dishes with ECMatrix-221 at 0.25 μg/cm² (for example, for one well of a 6-well plate add 1 mL of the 2.5 μg/mL working solution).
3. Incubate for 1 hour at 37°C, 3 hours at room temperate or overnight at 4°C.
4. Before use, remove remaining fluid from the coated surface (do not rinse).
5. Detach cells into small clumps using Accutase.
6. Plate the cells at desired density.
Note: Do not allow the plates to dry, briefly spin down all liquids in the tube before use, avoid repeated freeze-thaw cycles.
Analysis Note
Purity (SDS-Page): > 95%
Endotoxin Test: = 750 EU/mg
Mycoplasma Test: Negative
Sterility Test: Negative
Integrin Binding Assay (kDa): = 10 nM
Other Notes
6 X 175 μg ECMatrix-221 E8 Laminin Substrate (0.5 mg/mL in PBS). Expressed in CHO-S cells.
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
