HCE-2 cells were suspended and plated in pretreated 24-well plates (approximately 4 × 104 cells/cm2). Once they reached about 70–80% of confluence, the medium was removed and the cells were exposed for 5 h with hyperosmotic (450 mOsM) medium, obtained by sodium chloride (NaCl, 69 mM) addition [28 (link)]. The medium’s osmolarity was measured by the Advanced® Model 3320 Micro-Osmometer (Advanced Instruments, Norwood, OH, USA). Hyperosmolar cells were treated with TQ- (5 µM) free or formulated. Gene expression, protein levels and MitoSOX assay were performed after the treatments.
Advanced model 3320 micro osmometer
Manufactured by Advanced Instruments
Sourced in United States
The Advanced® Model 3320 Micro-Osmometer is a laboratory instrument designed to measure the osmolality of small sample volumes. It utilizes the freezing point depression method to determine the osmolality of a given sample. The instrument is capable of measuring osmolality within a range of 0 to 2000 mOsm/kg H2O, with a minimum sample volume requirement of 20 microliters.
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Lab products found in correlation
7 protocols using advanced model 3320 micro osmometer
1
Hyperosmotic Stress Response in HCE-2 Cells
2
Measurement of Urinary and Serum Markers
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Urinary osmolality was measured using an Advanced® Model 3320 Micro-Osmometer (Advanced instruments Inc., Norwood, MA, USA). A colorimetric assay was used to determine serum and urine Ca2+ concentrations as described previously [22 (link)]. Ca2+ measurements were verified using an internal control, which was a commercial serum standard (Precinorm U, Roche, Basel, Switzerland). Serum and urinary phosphate (Pi) concentrations were determined by in-hospital services using automatic biochemical analyzers. Serum PTH levels were determined using the mouse PTH 1–84 ELISA kit (Immunotopics international, San Clemente, CA, USA) and serum Fibroblast Growth Factor (FGF)23 levels were determined with the mouse C-terminal FGF23 ELISA assay (Immunotopics International).
3
Controlled Synthesis of Hyperbranched Polyglycerols
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Example 6
HPG polymer (0.5, 1, 3.5 kDa) was synthesized by anionic ring opening multi-branching polymerization of glycidol as described previously (Sunder, A., Hanselmann, R., Frey, H., Mulhaupt, R., 1999. Controlled synthesis of hyperbranched polyglycerols by ring-opening multibranching polymerization. Macromolecules 32, 4240-4246). The molecular characteristics of the polymer were determined by gel permeation chromatography and proton nuclear magnetic resonance spectroscopy. HPG was purified by dialysis against MilliQ filtered water and lyophilized. HPG-based preservation solutions were prepared by dissolving HPG (3%, w/v) in a solution containing: 100 mM lactobionic acid, 100 mM potassium hydroxide (KOH), 25 mM potassium dihydrogen phosphate (KH2PO4), 5 mM magnesium sulfate (MgSO4), 5 mM adenosine, 3 mM glutathione, and 1 mM allopurinol, the same composition as in Viaspan™ UW solution (UW solution, DuPont Canada, Mississauga, ON, Canada) omitting 30 mM raffinose and 5% HES. The pH of HPG preservation solution was adjusted to 7.4 using NaOH/HCl at 22° C., and its osmolality (˜320 mOsm/kg) was determined using Advanced® Model 3320 Micro-Osmometer (Advanced Instruments, Inc., Norwood, Mass., USA) in the Vancouver Coastal Health Regional Laboratory Medicine (Vancouver, BC, Canada).
4
Assessing Kidney Injury Biomarkers in Exercise
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At baseline, after 30 min and directly after completing the submaximal exercise test, all subjects provided a urine sample to examine fluid balance and kidney responses. Urinary cystatin C concentration was measured using the nephelometric method (Behring Nephelometer II, Siemens Healthcare, Den Haag, The Netherlands). The urinary creatinine concentration was measured using an enzymatic assay (Cobas C6000, Roche Diagnostics, Indianapolis, USA). Urine osmolality was examined using an osmometer (Advanced Model 3320 Micro‐Osmometer, Osmometer, Advanced Instruments, Norwood, USA).
In order to determine kidney injury in response to exercise, we measured urinary concentrations of KIM1 and NGAL (both monomeric and dimeric) in duplicate using the previously described sandwich ELISA assay (E‐EL‐H0186 and E‐EL‐H0096, Elabscience Biotechnology, Wuhan, China) (Han et al.2002 ; van Timmeren et al. 2007 ). Furthermore, uKIM1 and uNGAL concentrations were corrected for urinary cystatin C, creatinine, and osmolality. The corrected uKIM1 and uNGAL data were calculated by dividing the individual uncorrected data by the corresponding cystatin C levels, creatinine levels, and the urine osmolality. Additionally, urinary albumin (uAlbumin) and glucose (uGlucose) concentrations were measured, and corrected as well, to examine the effects of acute exercise and dehydration on acute kidney injury.
In order to determine kidney injury in response to exercise, we measured urinary concentrations of KIM1 and NGAL (both monomeric and dimeric) in duplicate using the previously described sandwich ELISA assay (E‐EL‐H0186 and E‐EL‐H0096, Elabscience Biotechnology, Wuhan, China) (Han et al.
5
Urine Analysis in Townes Mice
6
Osmolality Measurement of FB-PεCL-NPs
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The osmolality of ~50 µL of each FB-PεCL-NPs formulation was measured by means of Advanced® Model 3320 Micro-Osmometer (Advanced® Instruments, Inc., Norwood, MA, USA).
7
Preparation and Characterization of HPGS
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The HPGS was prepared by dissolving 1 kDa HPG polymer (3%, wt/vol) in a chemical composition similar to Belzer UWS (Bridge to Life Ltd, Columbia, SC) (Table S1 ) as described previously.21 , 22 The pH of the HPGS was measured and adjusted to 7.2–7.4 with NaOH/HCl solutions by using a Fischer Scientific Accumet pH meter (Fisher scientific, Ottawa, ON, Canada), and its osmolality (mOsm/kg) was determined by using the Advanced® Model 3,320 Micro‐Osmometer (Advanced Instruments, Inc., Norwood, MA) in the Vancouver Coastal Health Regional Laboratory Medicine (Vancouver, BC, Canada).
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