Rtn3 and GAPDH Protein Analysis

The lung tissue was lysed by radio-immunoprecipitation assay buffer (Thermo Fisher Scientific) supplemented with protease inhibitors, after which protein level was evaluated by a bicinchoninic acid protein detection kit (Thermo Fisher Scientific). The protein was analyzed with sodium dodecyl sulfate polyacrylamide gel electrophoresis. Subsequently, the protein sample on the polyvinylidene fluoride membrane was blocked, combined with primary antibodies Rtn3 and GAPDH (both from Santa Cruz Biotechnology, CA, USA), and with peroxidase-labeled secondary antibody (Abcam, CA, USA). The protein bands were analyzed with Odyssey 3.0 [23 (link)].

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Yang S., Liu F, & Wang D. (2022). Long noncoding RNA Kcnq1ot1 prompts lipopolysaccharide-induced acute lung injury by microRNA-7a-5p/Rtn3 axis. European Journal of Medical Research, 27, 46.

Publication 2022

radio-immunoprecipitation assay buffer bicinchoninic acid protein detection kit GAPDH peroxidase-labeled secondary antibody

Antibodies Antibody Assay Bicinchoninic acid Buffer Gapdh Immunoprecipitation Lung Membrane protein Peroxidase Polyvinylidene fluoride Protease inhibitors Protein Protein a Sodium dodecyl sulfate polyacrylamide gel electrophoresis Tissue

Corresponding Organization : Daqing City People's Hospital

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Variable analysis

independent variables

Radio-immunoprecipitation assay buffer (Thermo Fisher Scientific) supplemented with protease inhibitors

dependent variables

Protein level
Protein bands

control variables

Sodium dodecyl sulfate polyacrylamide gel electrophoresis
Polyvinylidene fluoride membrane
Primary antibodies Rtn3 and GAPDH (both from Santa Cruz Biotechnology, CA, USA)
Peroxidase-labeled secondary antibody (Abcam, CA, USA)
Odyssey 3.0 for protein band analysis

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

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