Total protein from BMDM, neutrophils, lymphocytes and mentioned tissues was extracted in ice cold RIPA buffer (C-9806S, Cell Signaling Tech. Beverly, Massachusetts) containing protease inhibitors (P8340, Sigma Aldrich St. Louis, Missouri). Proteins were detected by immunoblotting using standard techniques. Antibodies that were used: caspase-11 (sc-374615), GAPDH (sc-47724), β-ACTIN (sc-47778), Pol II (sc-9001), GCN5 (sc-20698) from Santa Cruz; caspase-1(AG-20B-0042) from adipogen; anti-COMMD10 antibody (GTX121488) from Genetex, pP65 (3033) and P65 (6956) from Cell Signaling. Blots were incubated with HRP-conjugated secondary antibodies, and subjected to chemiluminescent detection using the MicroChemi imaging system (DNR Bio-Imaging Systems, Israel). P65 subcellular fractionation was performed using NE-PER nuclear/cytoplasmic extraction kit (78835, Thermo scientific, Paisley, UK) per manufacturer's instructions. Equivalent protein amounts were loaded for both nuclear and cytoplasmic fractions.
Microchemi imaging system
Manufactured by DNR Bio-Imaging Systems
Sourced in Israel
The MicroChemi imaging system is a compact and versatile imaging device designed for a wide range of applications in life science research. It utilizes advanced imaging technologies to capture high-quality images of various samples, such as Western blots, gels, and membranes. The MicroChemi system enables users to perform quantitative analysis and documentation of their experimental results.
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Lab products found in correlation
3 protocols using microchemi imaging system
1
Immunoblotting Protein Extraction Protocol
2
Liver Protein Extraction and Immunoblotting
3
Western Blot Analysis of Phosphorylated ERK
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Cartilage explants (5-mm diameter discs made with a skin punch biopsy tool) incubated for 48 h in serum-free medium and then cut into smaller pieces, or cultured chondrocytes, were lysed with icecold cell-lysis buffer (50 mM TriseHCl, pH 7.2; 150 mM NaCl; 0.5% (v/v) Tritonx-100; 5 mM EDTA) supplemented with protease inhibitor cocktail (Roche, UK). Lysates were resolved by sodium dodecyl sulfate polyacrylamide electrophoresis, transferred to nitrocellulose membranes and subsequently probed using the antibodies described above. The signal was visualized with enhanced chemiluminescence and photographed with a MicroChemi imaging system (DNR Bio Imaging systems ltd). The intensity of the bands was quantified by densitometry with the NIH Image J software (1.49n; National Institutes of Health). The relative phosphorylation levels were calculated as the ratio of phospho-ERK to total ERK (tERK) signal intensities and were normalized to the control (no stimulus).
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