2 chloro l phenylalanine

Glyoxylic acid (50% in water) was purchased from TCI (Tokyo, Japan). The iTRAQ kit was purchased from AB SCIEX (113 Foster City, CA, USA). Methanol and acetonitrile were purchased from Merck (Darmstadt, Germany). Ultrapure water was prepared using a Milli-Q water purification system (Millipore Corp., Billerica, MA, USA). Formic acid, ammonium formate, and 2-chloro-L-phenylalanine were obtained from Sigma-Aldrich (St. Louis, MO, USA).

Methanol (HPLC grade) and acetonitrile (HPLC grade) were purchased from Merck (Darmstadt, Germany). Formic acid (FA) (HPLC grade) was purchased from the Fluka Chemical Corp (Buchs, Switzerland). 2-Chloro-L-phenylalanine (as an internal standard) was purchased from Sigma-Aldrich Company (St Louis, MO, USA). Hydroxyproline (HLP) (99% purity) was purchased from GL Biochem Ltd. (Shanghai, China). Sodium carboxymethylcellulose (CMC-Na) was purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). Chromatography Mass prepared by the Milli-Q system (Millipore, Bedford, MA, USA) with ultrapure water. Kidney injury molecule-1 (KIM-1) ELISA kit was purchased from Wuhan Uscn Life Science, Inc. (Wuhan, China). Von Kossa kit was purchased from GENMED Scientific Inc. (Shanghai, China). Hydroxyproline (300 mg/mL) was dissolved into 0.5% CMC-Na and stored at 4 °C.

For LC-MS, methanol (HPLC grade), acetonitrile (HPLC grade), formic acid (HPLC grade), and 2-chloro-L-phenylalanine (as an internal standard) were purchased from Sigma. (Saint Louis, MO, USA). Ultrapure water was produced using the Milli-Q system (Millipore, Bedford, MA, USA).
Animal treatment took corn oil (GLBIO, Montclair, CA, USA) as well as estradiol (GLBIO, Montclair, CA, USA) into application. The low limb samples used for micro computed tomography (Micro-CT) and histopathology examinations were preserved applying 80% paraformaldehyde stationary liquid (Labgic Technology, Beijing, China).
In the bone tissue decalcification process, EDTA-2Na and sodium hydroxide were purchased from Sinopharm Chemical Reagent Co., Ltd., Shanghai, China. For hematoxylin-eosin (HE) staining, tartrate-resistant acid phosphatase (TRAP) staining and alkaline phosphatase (ALP) staining, anhydrous ethanol was purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). The environmentally friendly dewaxing agent, transparent agent and sealed tablet were purchased from Tonsen tech, Wuhan, China. The histopathological staining kits were purchased from Biossci (Beijing, China) for HE staining (model BP092), TRAP staining (model BP088), and ALP (Model BP090).

Ultrapure water was produced by a Ming Che D24 UV water purification system (Merck Millipore, Burlington, USA). MeOH, ACN, NH₄OAc, and NH₄OH (CNW Technologies, Düsseldorf, Germany) were all MS grade. 2-Chloro-l-phenylalanine (Sigma, St. Louis, MO, USA) was used as an internal standard. HEPES, SDS, TFA, triethylammonium bicarbonate (TEAB), ammonium formate, Columbia agar, MacConkey agar, and Chocolate agar were obtained from Sigma (St. Louis, MO, USA). Dithiothreitol (DTT), iodoacetamide (IAA), and trypsin (sequencing grade) were obtained from Promega Life Sciences (Madison, USA). Urea was provided by GibcoBRL (MD, USA). EDTA and PMSF were obtained from Amesco (USA). Ethanol, formic acid, acetone, BCA protein quantification kit, TMT 6 standard kit, Acclaim PepMap C18 Column (100 μm × 20 mm, 75 μm × 250 mm) were purchased from Thermo Fisher Scientific (Waltham, MA, USA). TEAB and 25% ammonia were obtained from Santa Cruz (CA, USA). Sep Pak C₁₈ cartridges (1 cc, 100 mg) and the Acquity UPLC®BEH C18 column (50 mm × 2.1 mm, 1.7 μm) were obtained from Waters (Milford, USA). The Luria–Bertani (LB) broth and LB agar were purchased from Becton Dickinson (Sparks, MD, USA). The lysis buffer for protein analysis was obtained from Roche Ltd (Basel, Switzerland). All other reagents were of ACS reagent grade.

Methanol (HPLC grade), acetonitrile (HPLC grade), formic acid (HPLC grade), and 2-chloro-L-phenylalanine (as an internal standard) were purchased from Sigma. (Saint Louis, USA). Ultrapure water was produced using the Milli-Q system (Millipore, Bedford, MA, USA). Hematoxylin-eosin staining kit was purchased from Shanghai Ailex Technology Co., Ltd.BDE-3 (CAS 101-55-3, 99% purity) was purchased from Sigma-Aldrich (St. Louis, MO).