Muscle tissue samples were disrupted in a homogenizer (Psychotron NS-310EII; Microtec Co, Ltd, Chiba, Japan), and the protein concentrations were determined by the Bio-Rad protein assay (Bio-Rad Life Science, Hercules, CA). The relative expression of cytokines and chemokines in muscle lysates was quantified using the Proteome Profiler Array (Mouse Cytokine Array, Panel A; R&D Systems Inc., Minneapolis, MN). The array was performed according to the manufacturer's specifications with 100 mg of total protein. To achieve maximum assay intensity, the blots were incubated overnight. The chemoluminescence signal was generated by incubation with the Super Signal West Femto Chemiluminescent Kit (Thermo Scientific Pierce, Rockford, IL) for 5 minutes. Images were captured using Image Quant LAS 4000 coupled with Image Quant TL software version 1.51h (GE Healthcare, Tokyo, Japan).
Image quant tl software version 1.51h
Manufactured by GE Healthcare
Sourced in Japan
Image Quant TL software version 1.51h is a tool for the analysis and quantification of images, particularly those generated from gel electrophoresis and Western blot experiments.
Automatically generated - may contain errors
2 protocols using image quant tl software version 1.51h
1
Cytokine and Chemokine Quantification in Muscle Tissue
2
Gastrocnemius Muscle Protein Analysis
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Proteins from gastrocnemius muscle were separated by SDS-PAGE (12% polyacrylamide) and electroblotted onto a nitrocellulose membrane. After blocking with 1% bovine serum albumin, membranes were incubated with the respective antibody at 1:500 to 1:100 dilutions as follows: rabbit anti-mouse C5a (250565; Abbiotech, San Diego, CA), rabbit anti-mouse CD68 (ab125212), rabbit anti-iNOS antibody (ab3523), rabbit anti-myeloperoxidase (ab65871), rabbit antitroponin T T1 (ab83907; Abcam), rabbit anti-CD163 (M-96), rabbit antieIL-1RA (H-110), rabbit anti-acetyl-CoA carboxylase (C83B10), rabbit antieAMP-activated protein kinase (AMPK)b1/2 (57C12), goat antieglyceraldehyde-3-phosphate dehydrogenase (V-18), goat anti-troponin T FS (C-18; Santa Cruz Biotechnology, Inc., Dallas, TX), rabbit anti-MYBPC1 (LS-C144479), and rabbit anti-MYBPC2 (LS-C101947; LSBio, Seattle, WA). Membranes were developed with the Super Signal West Femto Chemiluminescent Kit (Thermo Scientific Pierce), and images were captured using Image Quant LAS 4000 coupled with Image Quant TL software version 1.51h (GE Healthcare).
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