To determine protein concentrations, cells or tissues were lysed in RIPA lysis buffer containing a protease inhibitor cocktail (Roche). Proteins were separated by SDS-PAGE and transferred to nitrocellulose membranes. The membranes were incubated at 4°C overnight with one of the following primary antibodies: anti-β-actin (Cell Signaling Technology), anti-UCP1 (Santa Cruz Biotechnology), or anti-RIP140 (Abcam), followed by incubation with horseradish peroxidase–conjugated secondary antibodies. Signals were detected using ChemiDOCTMXRS+ and the Image LabTM system (Bio-Rad).
Image labtm system
Manufactured by Bio-Rad
Sourced in United States
The Image Lab system is a versatile imaging platform designed for life science laboratories. It enables capture, analysis, and documentation of a variety of sample types, including gels, blots, and chemiluminescent signals. The system provides accurate, high-quality imaging results.
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Lab products found in correlation
4 protocols using image labtm system
1
Protein Quantification via Western Blot
2
Western Blot Analysis Protocol
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WB analysis was performed as described previously [28 (link),29 (link)]. Briefly, cells were lysed with WB/IP lysis buffer (Beyotime Biotechnology, P0013, Shanghai, China) containing protease inhibitor cocktail (Roche, 11697498001, Germany). Cell extracts were subjected to 10% SDS-PAGE, transferred onto 0.45 μm PVDF membranes (Millipore, Darmstadt, Germany), and blocked with 5% non-fat milk at 4 °C overnight. The membrane was clipped and probed with a primary antibody at room temperature for 2 h. After washes with TBST, the membrane was incubated with horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (BOSTER, BA1054, Wuhan, China) or goat anti-mouse IgG (BOSTER, BA1051, Wuhan, China) at room temperature for 1 h. Protein bands were exposed in an Image LabTM System (BIO-RAD, Contra Costa County, CA, USA) after the addition of chemiluminescent substrate (Advansta, K-12045-D50, San Jose, CA, USA). The relative intensities of Western blots were quantified using Image J. A protein molecular weight marker was purchased from Bioscience (Double Helix, P12083, Shanghai, China).
3
Western Blot Analysis of Adipose Tissue
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Samples of adipose tissues in lysis buffer with phospho-stop and proteinase inhibitor was centrifuged at 12,000 g for 5 min supernatant, and then was collected. Lysates were mixed in the 5×SDS protein sample loading buffer and heated to 100°C for 10 min. Samples were run on a 10% Bis-Tris criterion gel (Bio-Rad Laboratory Inc., Hercules, CA) at 80 volts for 30 min electrophoretic transferred at 120 V for 45–60 min. Primary antibodies was incubated for 16 h at 4°C, washed three times for each 10 min and then for seconed antibodies for 1 h in room temperature. Signals were detected using the ChemiDOCTMXRS+ and the Image LabTM system (BIO-RAD, USA).
4
Western Blot Analysis of Adipose Tissue
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Adipose tissue lysates were mixed in the 5×SDS loading buffer and heated to 100°C for 10 min. Samples were run on a 12% Bis-Tris criterion gel (Bio-Rad, Hercules, CA, USA), incubated with primary antibodies overnight, and then incubated with the second anti-rabbit IgG (W4018, PROMEGA, USA) or anti-goat IgG. Signals were detected using the ChemiDOCTMXRS+ and the Image LabTM system (BIO-RAD, USA).
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