All content on this website, including dictionary, thesaurus, literature, geography, and other reference data is for informational purposes only. HEPES-buffered saline (HBS) solution: Prepare 2 × HBS stock by adding the following to 400 ml dH2O: 50 ml 1 M HEPES, 28 ml 5 M NaCl and 1.5 ml 0.5 M Na2HPO4. HEPES-buffered saline (HBS) solution: Prepare 2 × HBS stock by adding the following to 400 ml dH 2 O: 50 ml 1 M HEPES, 28 ml 5 M NaCl and 1.5 ml 0.5 M Na 2 HPO 4. Soluble VIP36 tetramer bound to high molecular weight high mannose-type glycans on the cell surfaces. In a polystyrene cuvette, add 815 μl HEPES buffer, 10 μl lead acetate, and 100 μl tissue extract (total volume 925 μl). 11.3). The HBS, CaCl2, and NaB solutions should all be sterile-filtered with 0.2 μM filters, aliquoted and stored at − 20 °C. Incubate on ice for 20 min. HeLaS3 cells (none), cells treated with 2 μg/ml kifunensine for 24 h (KIF), or kifunensine-treated cells subjected to endo H digestion (KIF/endo H) were incubated with 10 μg/ml PE-labeled soluble VIP36 tetramer (filled histogram) or PE-SA as a control (thin line) in the presence of 1 mM CaCl2, and then analyzed by flow cytometry. After washing twice with HBS, cells were suspended in 200 μl of HBS containing 1 μg/ml PI. Q-Sepharose Fast Flow (Amersham Pharmacia Biotech, Piscataway, NJ). Johanna Laukkanen, Seppo Ylä-Herttuala, in Methods in Enzymology, 2002, HEPES (5 mM) and 5 mM HEPES–20% (v/v) glycerol, Buffer A: 20 mM Tris-HCl (pH 8), 150 mM NaCl, 1 mM CaCl2, Buffer B: 20 mM Tris-HCl (pH 8), 150 mM NaCl, 1 mM CaCl2, BSA (2 mg/ml), 0.01% (w/v) NaN3, Western blot sample buffer: 62.5 mM Tris-HCl (pH 6.8), 10% (v/v) glycerol, 2% (w/v) SDS, 5% (v/v) 2-mercaptoethanol, 0.05% (w/v) bromphenol blue (in nonreducing buffer, 2-mercaptoethanol is replaced by deionized water), Oil red O: 60 ml of Oil red O and 40 ml of dextrin are combined and filtered through filter paper. Calculate the contribution of CBS to total H2S production from the assay in the presence of propargylglycine. Subtract the contribution of CBS from the total H2S production rate to determine the contribution of CSE. Place the cuvette in a cuvette holder for 5 min, at 37 °C maintained using a circulating water bath. Agar is melted and kept liquid in a 60 ° water bath and added to 37 ° medium containing reagents. Wash cells twice with HCO3 buffer. Genital herpes, often simply known as herpes, may have minimal symptoms or form blisters that break open and result in small ulcers. 2-Deoxyglucose (2-DOG), unlabeled and 14C-labeled, Lysis buffer: Triton X-100 lysis buffer (1% Triton X-100, 20 mM Tris, and 150 mM NaCl), Cytochalasin B: 0.96 mg/ml (2 mM) dissolved in ethanol and stored at − 20 °C, Bree K. Grillo-Hill, ... Diane L. Barber, in Methods in Cell Biology, 2014, Nigericin buffers of at least 2 known pH values, 1 μM BCECF working solution in either tissue culture media without FBS or HCO3 buffer. ScienceDirect ® is a registered trademark of Elsevier B.V. ScienceDirect ® is a registered trademark of Elsevier B.V. URL: https://www.sciencedirect.com/science/article/pii/S0076687910780113, URL: https://www.sciencedirect.com/science/article/pii/B9780123979452000019, URL: https://www.sciencedirect.com/science/article/pii/B9780123865090000119, URL: https://www.sciencedirect.com/science/article/pii/S007668790129106X, URL: https://www.sciencedirect.com/science/article/pii/S0076687902530597, URL: https://www.sciencedirect.com/science/article/pii/S0076687914000810, URL: https://www.sciencedirect.com/science/article/pii/S0076687915006126, URL: https://www.sciencedirect.com/science/article/pii/B9780123884480000279, URL: https://www.sciencedirect.com/science/article/pii/B9780128002803000049, URL: https://www.sciencedirect.com/science/article/pii/B9780124201385000239, Gene Transfer Vectors for Clinical Application, Christopher R. Logg, ... Noriyuki Kasahara, in, Regulators and Effectors of Small GTPases, Johanna Laukkanen, Seppo Ylä-Herttuala, in, Hydrogen Sulfide in Redox Biology, Part A, Martin Gustavsson, ... Tracy M. Handel, in, Imaging and Spectroscopic Analysis of Living Cells, Methods of Adipose Tissue Biology, Part B, Bree K. Grillo-Hill, ... Diane L. Barber, in. Krebs Ringer HEPES (KRH) buffer with 0.01% BSA, with or without 5 mM glucose, 200 nM adenosine, pH 7.4. Protease inhibitor cocktail tablets (Complete, Boehringer Mannheim, Indianapolis, IN, 1 tablet/50 ml) or a mixture of separate inhibitors [10 μM pepstatin, 5 μM aprotinin, 1 μg/ml tosyl-l-lysine chloromethyl ketone (TLCK), 10 μM leupeptin, 1 mM phenylmethylsulfonyl fluoride (PMSF), or 2 μg/ml 4-(2-aminoethyl)-benzenesulfonyl-fluoride (AEBSF)] can be used. Copyright © 2020 Elsevier B.V. or its licensors or contributors. Gibco HEPES (4- (2-hydroxyethyl)-1-piperazineethanesulfonic acid ) is a zwitterionic organic chemical buffering agent and is categorized as a "Good" buffer which derives from a set of buffers described by Dr. Norman Good and his colleagues in 1966 … Martin Gustavsson, ... Tracy M. Handel, in Methods in Enzymology, 2016, cOmplete EDTA-free protease inhibitor tablets (Roche, 5056489001), Rüdiger Rudolf, ... Marco Mongillo, in Methods in Enzymology, 2012, Heart perfusion solution: HEPES-buffered Tyrode solution (HE-Tyrode), 100% O2 gassed, Dye loading solution (if applicable): HE-Tyrode, Fluo-4AM (10 mM)/Pluronic F127 (10 mM), SP, Imaging solution: HE-Tyrode, BDM (10 mM), (±)-Blebbistatin (100 nM, Tocris, MI, USA), ViscOphtal (Winzer Pharma), artificial tear gel for objective immersion, Mi-Jeong Lee, Susan K. Fried, in Methods in Enzymology, 2014. The fluorescence of stained cells was measured using flow cytometry. 4.0 mg ml− 1 purified human CBS or purified human CSE in 100 mM HEPES, pH 7.4. These typically heal over two to four weeks. Caption: FIGURE 2: Fluorescence response of B (1 [micro]M) in THF: For the MB-AuNPs synthesis we used three concentrations of. Elizabeth Richey, Hongmin Qin, in Methods in Enzymology, 2013, Add double-distilled H2O to 500 ml, use KOH to adjust pH to 7.4, 1 mM DTT (add 1000 × stock solution right before use), 0.25 mM EGTA (0.5 ml of 0.5 M EGTA, pH 8.5 for 1 l), 10 mM HEPES (10 ml of 1 M HEPES for 100 ml), 5 mM MgSO4 (5 ml of 1 M MgSO4 for 100 ml), 5 mM EDTA (10 ml of 0.5 M EDTA, pH 8.0 for 100 ml), Christopher R. Logg, ... Noriyuki Kasahara, in Methods in Enzymology, 2012. Warm solutions to 37 °C prior to the experiment: Add 1 ml 1 μM BCECF in HCO3 buffer to cells.