Western Blot Analysis of Lung and Cell Samples

The method of western blot was performed as previously described (Pan et al., 2018 (link)). Briefly, lung tissues or RAW264.7 cells were lysed with RIPA lysis buffer with the protease inhibitor cocktail (Beyotime, Shanghai, China). Protein concentrations were determined by the BCA assay kit (Beyotime, Shanghai, China). Proteins were separated by SDS-PAGE and electrotransferred onto PVDF membranes. Protein bands were detected by Enhanced Chemiluminescent Substrate Kit (YEASEN, Shanghai, China) and imaged with Tanon 1600 (Tanon, Shanghai, China).

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Tian X., Guo M., Zhang X., Guo L., Lan N., Cheng Y., Han Y., Wang M., Peng Z., Zhou C, & Fan H. (2022). Strongylocentrotus nudus Eggs Polysaccharide Enhances Macrophage Phagocytosis Against E.coli Infection by TLR4/STAT3 Axis. Frontiers in Pharmacology, 13, 807440.

Publication 2022

protease inhibitor cocktail BCA assay kit Enhanced Chemiluminescent Substrate Kit

Assay Buffer Lung Membranes Protease inhibitor Protein Pvdf Raw264 7 cells Ripa Sds page Tissues Western blot

Corresponding Organization : Xiangnan University

Top 5 similar protocols

Variable analysis

independent variables

Lung tissues
RAW264.7 cells

dependent variables

Protein expression

control variables

RIPA lysis buffer with the protease inhibitor cocktail (Beyotime, Shanghai, China)
BCA assay kit (Beyotime, Shanghai, China)
SDS-PAGE
PVDF membranes
Enhanced Chemiluminescent Substrate Kit (YEASEN, Shanghai, China)
Tanon 1600 (Tanon, Shanghai, China)

controls

No positive or negative controls were explicitly mentioned.

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