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Custom Vector Construction> | ca 21 Materials and Methods: 22 Materials 23 24 25 26 27 The plasmid for SHH-barstar in a pET-21a(+) vector was purchased from Genscript (NJ, USA).... Expression, purification and labeling of barstar-SHH A plasmid was purchased from Genscript, which coded for a fusion protein with SHH at the N- terminus, followed by a spacer (EFPKPSTPPGSSGGAP)3, barstar and a pentahistidine tag at the C-terminus in a pET21a(+) plasmid. | Get A Quote |
Three-dimensional (3D) protein-patterned scaffolds provide a more biomimetic environment for cell culture than traditional two-dimensional surfaces, but simultaneous 3D protein patterning has proved difficult. We developed a method to spatially control the immobilization of different growth factors in distinct volumes in 3D hydrogels, and to specifically guide differentiation of stem/progenitor cells therein. Stem-cell differentiation factors sonic hedgehog (SHH) and ciliary neurotrophic factor (CNTF) were simultaneously immobilized using orthogonal physical binding pairs, barnase-barstar and streptavidin-biotin, respectively. Barnase and streptavidin were sequentially immobilized using two-photon chemi... More