Chloramine t trihydrate

Sulfuric acid, hydrochloric acid, formic acid, perchloric acid, sodium nitrate, sodium hydroxide, potassium hydroxide, potassium chloride, sodium sulfate anhydrous, potassium dihydrogen phosphate, chloramine T trihydrate, trichloroacetic acid, 2-thiobarbituric acid, ethylenediaminetetraacetic acid (EDTA), propylgallate, 4-(dimethylamino)benzaldehyde, petroleum ether (b.p. 40–60 °C), methanol, ethanol, hexane, acetonitrile, 2-propanol, chloroform, sodium methoxide solution, amino acid standard, L-tryptophan, hydroxyproline, L-2-aminobutyric acid, and 1,1,3,3-tetraehoxypropane were purchased from Sigma-Aldrich (Sigma-Aldrich Co., St. Louis, MO, USA).
Supelco 37 comp. FAME mix, linoleic acid methyl ester isomer mix were purchased from Supelco (Supelco Analytical, Bellefonte, PA, USA), trans FAME mix K110 from Grace (Grace, Deerfield, IL, USA).
AccQ Fluor reagent kit and AccQ Tag eluent A concentrate was purchased from Waters (Waters Corp., Miliford, MA, USA). Boric acid was purchased from AFT (Bratislava, Slovakia), Kjeltabs from Velf Scientifica (Velf Scientifica srl, Usmate, Italy).

In adaption to an essay from Reddy and Enwemeka [15 (link)] specimens were hydrolyzed in 2 mL reaction vessels with 6 M HCl (Carl Roth GmbH, Karlsruhe, Germany) for 20 h at 95 °C (Heating Block ThermoStat plus, Eppendorf, Hamburg, Germany). Due to the assay’s sensitivity regarding pH value, hydrolyzed samples were dried and dissolved again in DI water. Assay standard solutions in a range between 20 and 160 µg/mL were prepared dissolving L-hydroxy-proline (Sigma-Aldrich, Schnelldorf, Germany) in DI-water. For assay execution, 35 µL of each specimen and standard solution were transferred into a 96-well plate. 75 µL of chloramine-T reagent were added, which consisted of 1.27 g chloramine-T trihydrate (Sigma-Aldrich, Schnelldorf, Germany) in 20 mL n-propanol (Merck Millipore, Darmstadt, Germany), stocked up to 100 mL with acetate-citrate buffer per 100 mL solution. 75 µL of Ehrlich’s reagent (1.5 g 4-(dimethylamino)-benzaldehyde (Sigma-Aldrich, Schnelldorf, Germany) in 6.6 mL of n-propanol and 3.3 mL 70% perchloric acid (both Carl Roth GmbH, Karlsruhe, Germany) were added. Wells were sealed with Microseal^® ‘B’ PCR Plate Sealing Film (Bio-Rad, Watford, United Kingdom) followed by an incubation at 60 °C for 60 min. Coloration intensity measurement was performed by a photometer Infinite M200 PRO (Tecan, Crailsheim, Germany) at 570 nm.

Albumin, anti-vWF antibody, 5-bromo-2-deoxyuridine (BrdU), chloramine Ttrihydrate, corn oil, Ehrlich’s reagent, Evans blue dye, formamide, hematoxylin and eosin (H&E), Hexadecyl trimethylammonium bromide, hydroxyproline, O-dianisidine dihydrochloride, trichrome kit, and trichloroacetic acid were purchase from Sigma-Aldrich (St Louis, MO). LPS from E. coli 055: B5 was obtained from Santa Cruz Biotechnology, Inc. (Dallas, TX). RNeasy Mini kit, and DNase I were purchased from Qiagen USA (Germantown, MD). In Situ Cell Death Detection Kit, and SYBR Green master mix were obtained from Roche Diagnostics (Indianapolis, IN). Anti-BrdU antibody was purchased from BD Biosciences (San Jose, CA). DAPI, Alexa FITC-conjugated or Texas Red-conjugated secondary antibodies, and Trizol reagent were obtained from Thermo Fisher Scientific (Waltham, MA).

Chloramine T trihydrate, trans-4-Hydroxy-L-proline, 4-(dimethylamino) benzaldehyde and tamoxifen and other standard chemicals were purchased from Sigma (Taufkirchen, Germany).

Pepsin (Sigma-Aldrich, St. Louis, MO, USA), Hoechse33258 (Sigma-Aldrich, MA, USA), Papain (Sigma-Aldrich, MO, USA), HCl (SHOWA, Gyoda, Japan), NaOH (SHOWA, Japan), Triton X-100 (Sigma-Aldrich, MO, USA), Aprotinin (Sigma-Aldrich, MO, USA), RNase A (Sigma-Aldrich, MO, USA), Chloramine T trihydrate (Sigma-Aldrich, MO, USA), DNase I (Merck, Darmstadt, Germany), Citric acid (JTbaker, Radnor, PA, USA), Sodium acetate trihydrate (JTbaker, PA, USA), and Quant-iTTM PicoGreen dsDNA Reagent and Kits (Invitrogen, Waltham, MA, USA) were used.