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By Charles Streuli; Michael Grant

Top bench-top experimentalists from world wide current, in a confirmed layout, their state of the art equipment for probing the extracellular matrix (ECM), its mechanisms of meeting and the way it affects telephone habit. those rigorously certain biochemical, biophysical, genetic, molecular organic, and telephone organic ideas were optimized for the research of ECM constitution and meeting, the mechanisms of cell-ECM interactions, and the molecular genetics of ECM proteins and defects in human affliction, and convey to a much broader viewers quite a number hugely subtle equipment that experience hitherto been tricky even for a number of really good laboratories. state of the art and hugely useful, Extracellular Matrix Protocols deals cutting-edge ECM researchers a gold-standard choice of strong, comfortably reproducible tips on how to examine the extracellular matrix and the altered cell-ECM interactions that play so very important a task in human future health and sickness.

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Biol. Chem. 270, 9398–9406. 3. Fox, J. , Wiedemann, H. et al. (1991) Recombinant nidogen consists of three globular domains and mediates binding of laminin to collagen type IV. EMBO. J. 10, 3137–3146. 4. , Salzer, J. , Yurchenco, P. , and Tuomanen, E. I. (1997) Neural targeting of Mycobacterium leprae mediated by the G domain of the laminin-alpha2 chain. Cell 88, 811–821. 5. , O’Rear, J. , and Yurchenco, P. D. (1993) Cell and heparin binding in the distal long arm of laminin: identification of active and cryptic sites with recombinant and hybrid glycoprotein [published erratum appears in J.

Materials for Expression of Recombinant Human Type III Procollagen in Insect Cells 1. pVL1392 and p2Bac baculovirus expression vectors (Invitrogen, San Diego, CA). 2. Full-length cDNAs for the α- (12) and β- (13) subunits of human prolyl 4-hydroxylase and the proα1 chain of human type III procollagen [proα1(III)] (14). 3. Wizard Plus Maxiprep DNA purification system (Promega, Madison, WI). 4. Sf9 and High Five (H5) insect cells (Invitrogen). 5. TNM-FH medium (Sigma, St. Louis, MO) supplemented with 10% fetal bovine serum (FBS).

72, 325–398. 6. Kivirikko, K. I. and Myllyharju, J. (1998) Prolyl 4-hydroxylases and their protein disulfide isomerase subunit. Matrix Biol. 16, 357–368. 7. , and Kivirikko, K. I. (1992) Characterization of the human prolyl 4-hydroxylase tetramer and its multifunctional protein disulfide-isomerase subunit synthesized in a baculovirus expression system. Proc. Natl. Acad. Sci. USA, 89, 7467–7470. 8. , and Kivirikko, K. I. (1997) Assembly of human prolyl 4-hydroxylase and type III collagen in the yeast Pichia pastoris: formation of a stable enzyme tetramer requires coexpression with collagen and assembly of a stable collagen requires coexpression with prolyl 4-hydroxylase.

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