Polyclonal rabbit antigoat immunoglobulins hrp

The whole-cell lysate was prepared using SDS gel loading buffer. The lysate was separated by SDS–polyacrylamide gel electrophoresis (8% acrylamide) and transferred to PVDF membranes. Primary antibodies were incubated in TBS-T (0.05 M Tris-buffered saline and 0.05% Tween 20) with 10% FBS after blocking with 5% skim milk. EnVision+ System-HRP Labelled Polymer antimouse or antirabbit antibodies and polyclonal rabbit antigoat immunoglobulins/HRP (DAKO) were used as secondary antibodies. Detection was performed using a ChemiDoc MP System (Bio-Rad Laboratories) with HRP chemiluminescence substrate. Signal intensity was also measured using Image Lab Software.

4‐Methylumbelliferyl β‐D‐glucopyranoside, 4‐methylumbelliferyl N‐acetyl‐β‐D‐glucosaminide, sodium taurocholate hydrate, and ambroxol hydrochloride were purchased from Sigma‐Aldrich (St Louis, MO). Pierce BCA Protein Assay, Halt Protease Inhibitor Cocktail, Halt Phosphatase Inhibitor, Pierce ECL Western Blotting Substrate, and Power SYBR Green PCR Master Mix were purchased from Thermo Scientific (Waltham, MA). Luminata Forte Western HPR Substrate was purchased from Millipore (Billerica, MA). RNeasy Mini Kit was purchased from Qiagen (Hilden, Germany). Precision nanoScript 2 Reverse Transcription kit (RT‐nano‐Script2) was purchased from Primerdesign (Chandler's Ford, UK). Anti–α‐synuclein antibody (4D6; ab1903), anti–α‐synuclein (phospho S129) antibody (EP1536Y; ab51253), anti–mitochondrial transcription factor A (TFAM) antibody (ab131607), and anti–transcription factor EB (TFEB) antibody ‐ ChIP grade (ab2636) were purchased from Abcam Biochemicals (Cambridge, UK). Polyclonal swine antirabbit immunoglobulins/horseradish peroxidase (HRP), polyclonal goat antimouse immunoglobulins/HRP, and polyclonal rabbit antigoat immunoglobulins/HRP were purchased from Dako (Glostrup, Denmark).

The dentin sialoprotein was evaluated through immunocytochemistry. Cells were prepared and incubated with a primary antibody (DSP (M-20) Antibody, Santa Cruz Biotechnology, Europe, 1:100) and a peroxidase-conjugated secondary antibody (Polyclonal Rabbit Anti-Goat immunoglobulins/HRP, Dako, Denmark, 1:100). Images were obtained on a Moticam 1080 optical microscope (Microscope Central, Bustleton Pike, Feasterville, PA, USA).