Original data of WB (Pi lab.2025.Mar)

Description

PT109B Rapidly Converts Astrocytes to Dopaminergic Neurons and Alleviates Parkinson's Disease in Mice.

Steps to reproduce

Cells were lysed using RIPA lysis buffer (containing protease inhibitors), and the resulting lysates were centrifuged to remove cellular debris. Protein concentration was determined using a bicinchoninic acid (BCA) Protein Assay Kit (ThermoFisher, 23225) following the manufacturer’s instructions. Based on protein concentration quantified by BCA, 30 µg of protein per well was loaded onto the gel. Electrophoresis was performed at a constant voltage of 80 - 120V. The membrane was blocked with 5% bovine serum albumin (BSA) or 5% non-fat milk in Tris-buffered saline with 0.1% Tween-20 (TBST) for 2-3 hours at room temperature to prevent non-specific binding. The blocked membrane was incubated with the primary antibody diluted in TBST at 4°C overnight and a horseradish peroxidase (HRP)-conjugated secondary antibody at room temperature for 1 hour. Protein bands were visualized using a cold charge-coupled device (CCD) imaging system after exposure to enhanced chemiluminescence (ECL) substrate.

Institutions

• Sun Yat-Sen University

  Guangdong, Guangzhou

Categories

Funders

• Shenzhen Municipal Science and Technology Innovation Council

  Guangdong, China

  Grant ID: No. 202111233000079

Licence