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Proc. Natl. Acad. Sci. USA 99 (231), 15072–15077. Proliferative action of mast-cell tryptase is mediated by PAR2, COX2, prostaglandins, and PPAR_: Possible relevance to human fibrotic disorders. 2002

Frungieri, M.B., Weidinger, S., Meineke, V., Kohn, F.M. and Mayerhofer, A.

Notes: Tryptase treated human fetal foreskin fibroblast cells (HFFF2s) showed a rapid increase in protigladin, COX2 and PAR2 mRNA, and protein levels. For these experiments 12, 120, or 1,200 milliunits/ml tryptase were applied to HFFF2s in culture.  Cells were then analyzed by various means for protein and mRNA levels. Tryptase was also shown to induce proliferation of HFFFs in culture. (2747)

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Biotechnol. Appl. Biochem. 28, 125-131. Recombinant human mast cell tryptase beta: stable expression in Pichia pastoris and purification of fully active enzyme. 1998

Niles, A.L., Maffitt, M., Haak-Frendscho, M., Wheeless, C.J., and Johnson, D.A.

Notes: Human mast cell tryptase was expressed and purified from Pichia pastoris by hydrophobic interaction chromatography followed by affinity chromatography on immobilized heparin. A faint band at 33Da and diffuse bands at 34.2, 35.9 and 50 kDa were observed on SDS/PAGE due to differences in the post-translational glycosylation of asparagine residues. Expression patterns of human tryptase were analysed by separating proteins by 10% SDS/PAGE , transferring to nitrocellulose membrane, and immunoblotting with the Anti-Human Tryptase mAb Biotin at 100 ng/ml in TBST. The blots were incubated with a 1:5000 dilution of Promega's Anti-Mouse IgG (H+L), AP Conjugate and developed with the Western Blue® Stabilized Substrate for Alkaline Phosphatase. (2451)

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