A total of 3 OLP-MFs and 3 NFs were selected for Simple Western analysis. Total protein expression of α-SMA and IL-6 were assessed using an automated capillary electrophoresis-sized-based Simple Western system using a Wes machine (ProteinSimple). Simple Western is a gel-free, blot-free, capillary-based, automated protein immunodetection system that automates all the steps following sample preparation, including sample loading, size-based protein separation, immunoprobing, washing, detection and data analysis (16 (link),17 (link)). All procedures were performed using the manufacturer's reagents and according to protocol. Data were analyzed with Compass software (version 4.0; ProteinSimple). Anti-α smooth muscle Actin antibody (1:2,000; cat. no. ab5694; Abcam), anti-IL6 antibody (1:2,000; cat. no. ab6672; Abcam) and GAPDH (clone 6C5; 1:5,000; cat. no. MAB374-AF647; Chemicon International; Thermo Fisher Scientific, Inc.) were used, with GAPDH being used as the loading control.
Protocol detail
Simple Western Analysis of α-SMA and IL-6
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Actin
Af647
Anti antibody
Capillary
Capillary electrophoresis
Clone
Gapdh
Protein
Smooth muscle
Corresponding Organization :
Other organizations : Southern Medical University
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Variable analysis
independent variables
- Selection of 3 OLP-MFs and 3 NFs
- Total protein expression of α-SMA
- Total protein expression of IL-6
- GAPDH (clone 6C5; 1:5,000; cat. no. MAB374-AF647; Chemicon International; Thermo Fisher Scientific, Inc.) was used as the loading control
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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