Tgf beta activation protocol

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Advertisement Hide. Protocol First Online: 25 May This is a preview of subscription content, log in to check access. Front Pharmacol J Biol Chem — Proc Am Thorac Soc — Schultz-Cherry S, Murphy-Ullrich JE Thrombospondin causes activation of latent transforming growth factor-beta secreted by endothelial cells by a novel mechanism. J Cell Biol — J Immunol — Nat Med — PLoS One e Chest — CrossRef Google Scholar. It gets diluted with dH 2 O.

Add 35ul of lysis buffer to each well. Lyse cells for 15 minutes RT on a shaker. At this point you can cover samples air-tight with Parafilm and store them at C until ready to assay. Get opaque well assay plate. Make sure it is clean! Transfer 30 ul of each sample to the new plate. Ahead of time, make reaction buffer. This is prepared using premade frozen aliquots of luciferin, ATP and buffer mix- these aliquots are combined and water added to final volume of 10 ml.

Reaction buffer should be RT before it is used. Set up luminometar for the assay. Put assay plate into holder. Run assay. After reading is done, flush the tubings with water, then empty the tubings and turn off the machine. Buffer stock Reagent Stock conc Final conc.