iMatrix-221 Recombinant Human Laminin-221 E8 Fragment Protein

- Overview
  - iMatrix-221 is a recombinant human laminin-221 E8 fragment protein expressed in Chinese Hamster Ovary (CHO)-S cells. iMatrix-221 contains the integrin-binding site of the laminin-221 molecule. iMatrix-221 is a useful cell culture substrate for proliferation and differentiation of cardiomyocytes and skeletal muscule cells. iMatrix-221 is also useful for the culture of other cells adhering to laminin-221.
    
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    More Details
    
    Recombinant Laminin Fragments
- Properties
  - Source
    
    CHO-S cells
    
    Type
    
    Recombinant Proteins
    
    Storage
    
    Store at 2°C to 15°C, protect from light.Shelf life is 2 years.
    
    Concentration
    
    0.5 mg/mL
    
    Molecular Weight
    
    150 kDa
    
    Reconstitution
    
    Recombinant human laminin-221 E8 fragment protein in PBS(-)
    
    Activity
    
    The dissociation constant (Kd) for the binding with integrin α7X2β1 is 10 nM or less.
    
    * For research use only. Not intended for human use.
- Applications
  - Application
    
    Cell culture
    
    Application Description
    
    Substrate for purification and maintenance of cardiomyocytes
    
    By the following method, iMatrix-221 can be coated onto a culture vessel. The optimum coating density may differ by cell-type, cell-line, medium selected, or purpose. Insufficient coating density may result in the detachment of cells and varied cell conditions while the excessive coating density may lead to difficulty in detaching cells for passage.
    Determine the optimal coating density. 0.5 μg/cm2 is a standard but test between 0.1 and 1.5 μg/cm2.
    1) Dilute iMatrix-221 with PBS(-). Use the diluted iMatrix-221 immediately. To coat with 0.5 μg/cm2 onto a 6-well plate with 9.6 cm2/well, dilute 9.6 μL of iMatrix-221 with 2 mL of PBS(-) per well.
    2) Place the diluted iMatrix-221 into a culture vessel and incubate either at 37°C for 1 h, or at room temperature for 3 h, or at 4°C overnight.
    3) Aspirate the coating solution. Then, immediately seed your cells. Do not allow the coated surface to dry.
    *If you face difficulties in detaching cells for passage, re-adjust the conditions (e.g., reduce the coating density).
- Reference
  - 1.Oliviero P. et.al., Expression of laminin α2 chain during normal and pathological growth of myocardium in rat and human. Cardiovascular Research 46, 346-355 (2000).
    2.Ja K.P. Myu Mia et.al., PSC-derived human cardiac progenitor cells improve ventricular remodeling via angiogenesis and interstitial networking of infarcted myocardium. J Cell Mol Med 20(2), 323-332 (2016).
    3.Unpublished, Osaka University (2018).
    4.Miyazaki T. et al. Laminin E8 fragments support efficient adhesion and expansion of dissociated human pluripotent stem cells. Nature Communications 3: 1236 (2012).
    5.Taniguchi Y. et al., The C-terminal region of laminin β-chains modulates the integrin-binding affinities of laminins. J. Biol. Chem.284 (12): 7820-31 (2009).
    6.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 (15): 11144-54 (2017).