Carboxymethyl cellulose sodium salt

The 10 ml of 35% hydrogen peroxide (Wako Pure Chemical), 5 ml of deionized water, and 0.35 g of carboxymethyl cellulose sodium salt (Wako Pure Chemical) were mixed. The carboxymethyl cellulose sodium salt was used as thickener to increase the viscosity of bleaching gel. Then each of sodium hydroxide (NaOH, Wako Pure Chemical), sodium bicarbonate (NaHCO₃, Wako Pure Chemical), sodium carbonate (Na₂CO₃, Wako Pure Chemical), potassium hydroxide (KOH, Wako Pure Chemical), potassium hydrogen carbonate (KHCO₃, Wako Pure Chemical), or potassium carbonate (K₂CO₃,Wako Pure Chemical) was added as a pH conditioner, and pH of the solution was adjusted as 7.0 (Table 1). Final concentration of hydrogen peroxide in the bleaching materials was 23.0%.

Atropine sulfate hydrate (special grade), acetonitrile (LC-MS grade), trifluoroacetic acid (HPLC grade), water (LC-MS grade), 2-methylbutane (isopentane), and carboxymethyl cellulose sodium salt (CMC) were purchased from FUJIFILM Wako Pure Chemical Corporation (Osaka, Japan). Acetone was purchased from Nacalai Tesque, Inc. (Kyoto, Japan). Hematoxylin and eosin were purchased from Sakura Finetec Japan Co. Ltd. (Tokyo, Japan). CHCA (α-cyano-4-hydroxycinnamic acid) was purchased from Bruker Daltonik GmbH (Bremen, Germany). The 1% atropine ophthalmic solution was obtained from Nihon Tenganyaku Kenkyusho Co. Ltd. (Nitten, Nagoya, Japan). Somnopentyl (pentobarbital sodium) was obtained from Kyoritsu Seiyaku Corporation (Tokyo, Japan).

Fluorescein Dolichos biflorus agglutinin (DBA; catalog no.: FL-1031) was purchased from Vector Laboratories (CA). Anti-ATP6V0A4 rabbit IgG (catalog no.: ab204737) and anti-lysosome-associated membrane protein 2 (LAMP2) rat IgG (catalog no.: ab13524) were purchased from Abcam (Cambridge, UK). Carboxymethyl cellulose sodium salt and isoflurane were purchased from Wako (Osaka, Japan). Alexa Fluor 488- and 594-conjugated anti-rabbit IgG were purchased from Thermo Fisher Scientific (Waltham, MA). Anti-calbindin D-28K rabbit IgG (catalog no.: AB1778; Research Resource Identifier: AB_2068336) was purchased from Merck Millipore (Burlington, MA). 9-Acridinylamine (9AA), galactocerebrosides from bovine brain (catalog no.: C4905), and α-cyano-4-hydroxycinnamic acid (CHCA; catalog no.: C2020) were purchased from Merck KGaA (Darmstadt, Germany). Hoechst 33258 was purchased from Nacalai Tesque (Kyoto, Japan). RNeasy Mini Kit and Rotor-Gene platform were purchased from Qiagen (Venlo, The Netherlands). ReverTra Ace quantitative PCR (qPCR) RT master mix with genomic DNA remover and Thunderbird qPCR Mix were purchased from TOYOBO (Osaka, Japan).