Cytokine/chemokine analyses from tumors of humanized mice were conducted using the Bio-Plex Pro™ Human Chemokine Panel, 40-Plex (Bio-Rad Laboratories AG, Cressier, Switzerland, Cat No.: 171AK99MR2). Small tumor fragments were snap frozen and whole protein was isolated in the presence of EASYpack Protease Inhibitor Cocktail (Roche; ref 5892970001) using the Precellys®24 Homogenizer and Bio-Plex® Cell Lysis Buffer following manufacturer instructions. Whole protein content was measured with BCA™ Protein Assay Kit (Thermo Scientific) before cytokine measurement was performed.
Precellys 24 homogenizer
Manufactured by Roche
The Precellys®24 Homogenizer is a high-performance benchtop homogenizer designed for the efficient disruption and lysis of a wide range of biological samples. It uses the principle of bead-beating technology to thoroughly homogenize samples, preparing them for subsequent analysis.
Automatically generated - may contain errors
Lab products found in correlation
Bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Bio plex pro human chemokine panel 40 plex, by Bio-Rad (1 mentions)
Easypack protease inhibitor cocktail, by Roche (1 mentions)
Magna lyser green beads, by Roche (1 mentions)
Pierce 660 nm protein assay, by Thermo Fisher Scientific (1 mentions)
Complete protease inhibitor cocktail, by Roche (1 mentions)
3 protocols using precellys 24 homogenizer
1
Cytokine/Chemokine Profiling of Tumor Xenografts
2
Ileum Membrane Protein Isolation
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Scrapings from mucosa of ileum were homogenized in a buffer containing (in mM) 200 mannitol, 80 HEPES, 41 KOH and protease inhibitors, pH 7.5. Homogenization was performed with MagNa Lyser Green Beads (Roche), in a Precellys 24 Homogenizer. Upon centrifugation at 800 rpm for 20 minutes at 4 °C, supernatants were further centrifuged at 41,000 rpm for 30 minutes at 4 °C, and pellets containing total membrane proteins were resuspended in the same buffer used for homogenization.
3
Synchronizing and Analyzing Worm Mutants
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xpc-1;csb-1 double mutants were synchronized by bleaching and fed for 3 hr or left under starvation. Six hours after UV (310 nm, Phillips UV6, Waldmann UV236B) or mock treatment, worms were collected in extraction buffer (50 mM HEPES KOH [pH 7.5], 300 mM NaCl, 1 mM EDTA, 1% [v/v] Triton X, 0.1% [w/v] sodium deoxycholate, 10% [v/v] glycerol, and complete protease inhibitor cocktail; Roche Diagnostics) and frozen in liquid nitrogen before resuspension in extraction buffer and homogenization with zirconia beads (four cycles, 6,000 × 2; 20 s; Precellys24 Homogenizer with Cryolys Cooling Unit). Supernatant was collected after 15 min of centrifugation at 4°C. The total protein concentration was measured using the Pierce 660 nm Protein assay (Thermo Fisher Scientific).
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