The total protein of the infiltrated N. benthamiana leaves and Eureka lemon leaves exhibiting transient expression was extracted using the Plant Total Protein Extraction Kit (Solarbio LifeSciences). Western blot analysis was performed as previously described (Du et al., 2022 (link)) and was repeated four times. The membranes were probed using monoclonal anti‐CYVCV CP, monoclonal anti‐GFP (Sigma), and anti‐His (Abmart) antibodies, followed by a horseradish peroxidase‐conjugated secondary antibody (Proteintech). Blots were visualized using a chemiluminescence detection kit (Everbright Inc.).
Monoclonal anti gfp
Manufactured by Merck Group
Monoclonal anti‐GFP is a laboratory reagent used to detect and quantify the presence of Green Fluorescent Protein (GFP) in biological samples. It is a highly specific antibody that binds to GFP, a widely used reporter protein in molecular biology and cell biology experiments.
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Lab products found in correlation
Horseradish peroxidase conjugated secondary antibody, by Proteintech (1 mentions)
Plant total protein extraction kit, by Solarbio (1 mentions)
Sds page gel, by Thermo Fisher Scientific (1 mentions)
Anti dpm1, by Thermo Fisher Scientific (1 mentions)
Chemidoc mp imaging system, by Bio-Rad (1 mentions)
2 protocols using monoclonal anti gfp
1
Protein Extraction and Western Blot Analysis
2
Yeast Whole Cell Protein Extraction
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Total protein extraction from yeast cells was performed by the NaOH-TCA lysis method (Ulrich and Davies, 2009) . Whole cell extracts and immunoprecipitated samples were separated on 10% or 4-12% precast SDS-PAGE gels (ThermoFisher Scientific). Proteins were further detected either by silver staining of the gel or by western-blot following transfer to PVDF membranes. The following validated antibodies were used: monoclonal anti-GFP (Sigma), 1:500; rabbit IgG-HRP polyclonal antibody (to detect protein-A-tagged proteins, Sigma), 1:5,000; anti-Dpm1 (ThermoFisher Scientific), 1:2,000. Specificity of anti-GFP antibodies was confirmed using untagged strains. Images were acquired using a ChemiDoc MP Imaging System (BIORAD).
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