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Chemiluminescence substrate system

Manufactured by Bio-Rad
Sourced in United States

The Chemiluminescence substrate system is a laboratory equipment product designed for the detection and quantification of target proteins in Western blot analysis. The system utilizes a chemiluminescent reaction to generate a luminescent signal that can be detected and measured using specialized imaging equipment. The core function of this system is to provide a sensitive and reliable method for the visualization and analysis of protein expression levels in biological samples.

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2 protocols using chemiluminescence substrate system

1

Western Blot Analysis of Lung Proteins

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Lung tissues or fibroblasts were homogenized in RIPA lysis buffer (Beyotime, Shanghai, China), to which two kinds of protease inhibitor cocktails (MCE, HY-K0022) had also been added. The proteins were separated on polyacrylamide gels and finally detected with a chemiluminescence substrate system (Bio–Rad Laboratories, CA, United States and Tanon, Shanghai, China) (Miao et al., 2020 (link)). The antibodies used for western blotting were as follows (Table 1).
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2

Lung Tissue Protein Expression Analysis

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The proteins from lung tissues have been extracted utilizing the RIPA Lysis Buffer containing protease inhibitor (GRF101, Epizyme) and phosphatase inhibitor cocktails (GRF102, Epizyme) while quantification performed utilizing a BCA Protein Assay Kit (ZJ101, Epizyme). Equivalent amounts of proteins have been electrophoresed on polyacrylamide gels and electrotransferred onto polyvinylidene fluoride (PVDF) membranes. The membranes were incubated for 2 h at room temperature by the primary antibodies which included ß-Tubulin antibody (HX 1829, huaxingbio), anti-FIZZ1 antibody (1:1,000, ab39626, Abcam) p38 mitogen-activated protein kinase (MAPK) Rabbit mAb (#8690, Cell Signaling Technology), Phospho-p38 MAPK (Thr180/Tyr182) Rabbit mAb (#4511, Cell Signaling Technology). Following a 40-min incubation at room temperature with secondary antibodies, the membranes were detected using a chemiluminescence substrate system (Bio-Rad Laboratories, CA, United States). The band intensities of proteins have been quantitated using Image J software.
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