hTRPV1-HEK cells were grown and maintained under standard cell culture conditions in high-glucose DMEM, supplemented with 10% FBS and 1% penicillin/streptomycin (Thermo Fisher Scientific, Waltham, MA). HEPES-Tyrode buffer solution was prepared with HEPES 0.025 mol, NaCl 0.140 mol, KCl 0.0027 mol, CaCl2 0.0018 mol, MgCl2 0.0005 mol, NaH2PO4 and D-glucose 0.05 mol (Sigma Aldrich). Fura-2AM (Invitrogen, Carlsbad, CA) was stored at −20°C and protected from light sources. A 20% (w/v) Pluronic F-127 (Sigma Aldrich) solution was prepared in-house in >99% DMSO and stored at room temperature for up to 1 hour before being transferred to cells at the time of assay. Stock solutions of the N-acyl ethanolamines (NAEs; Cayman Chemical) and capsaicin (Sigma Aldrich) were prepared in-house in 100% ethanol and stored at −80°C. Solutions of appropriate molarities were made immediately before experimentation, in >99% DMSO.
Hepes tyrode buffer
Manufactured by Merck Group
Sourced in United States
HEPES-Tyrode buffer is a laboratory solution commonly used as a physiological buffer. It is a balanced salt solution that helps maintain a stable pH and osmotic environment for cells and tissues in various in vitro experiments and applications.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using hepes tyrode buffer
1
Calcium Imaging of TRPV1 in HEK Cells
2
Calcium Mobilization in RBL-2H3 Cells
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RBL-2H3 cells (1 × 106/mL) were incubated with HEPES-Tyrode buffer (except for CaCl2 and MgCl2) (Sigma, USA) containing 1μ Ci45Ca2+/mL. Then, cells were treated with 50 ng/mL anti-DNP IgE for 6 h. Then, different concentrations of GLA were added for incubation for 30 min at 37°C followed by a challenge with 100 ng/mL DNP-HSA for 10 min. After cell lysis with 10% Triton X-100, radioactivity was determined by a scintillation β-counter (Liquid Scintillation Analyzer, Canberra Industries, USA).
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