Alveolar macrophages were washed with PBS and lysed with 1X RIPA buffer containing protease inhibitor cocktail (Thermo Scientific). Fifty micrograms of protein lysates were resolved by SDS-PAGE. Membranes were incubated with antibodies against FIZZ1 (Abcam), Arg (BD Biosciences) and CD206 (Abcam) antibodies. Signals were developed with Pierce ECL Western blot detection kit (Thermo Scientific, Rockford, IL).
Pierce ecl western blot detection kit
Manufactured by Thermo Fisher Scientific
Sourced in Japan
The Pierce ECL Western Blot Detection Kit is a chemiluminescent substrate used for the detection of proteins in Western blot analysis. The kit provides reagents necessary for the visualization of target proteins that have been separated by gel electrophoresis and transferred to a membrane.
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Lab products found in correlation
3 protocols using pierce ecl western blot detection kit
1
Macrophage Protein Expression Analysis
2
Western Blot Visualization Protocol
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Western blotting was performed as described in a previous paper.24 (link) The bound antibodies were visualized with a LAS-3000 imaging system (Fujifilm, Tokyo, Japan) using a Pierce ECL western blot detection kit (Thermo Scientific).
3
Quantifying Proteins from ZFAND3-Manipulated GSCs
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Total proteins from GSCs with ZFAND3 manipulations were extracted in RIPA lysis buffer and quantified using Bradford reagent (BioRad). After SDS-PAGE electrophoresis, proteins were transferred on nitrocellulose membrane (iBlot, Life Technologies). Primary antibodies were diluted in blocking solution (milk 5%) and incubated overnight at 4 °C (list of antibodies in Supplementary Table 3 ). After incubation with HRP-conjugated secondary antibodies, the chemiluminescent signal was recorded using the Pierce ECL Western blot detection kit (ThermoFisher), and the ImageQuant LAS4010 imaging station and software (GE Healthcare) and Image J.
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