Assessment of ricin preparations by SDS electrophoresis was carried out. The purity of the ricin preparations was assessed by SDS-PAGE using coomassie blue staining (GE Healthcare). Samples (1 mL at 5 or 10 mg/mL) were mixed with a 1:1 volume of sample buffer Tris-HCl (125 mM) pH to 6.8, SDS (10%, w/v), and glycerol (50%, v/v). The samples were heated at 95 °C for 5 min prior to loading 1 mL onto the gel (polyacrylamide gradient (4–14%); GE Healthcare). Proteins were separated for 40 min before coomassie blue staining for visualisation, using the manufacturer’s instructions (Calibrated Densitometer GS-800; Bio-Rad, Hemel Hempstead, UK). Densitometric scans were analysed using Quantity One software (Bio-Rad). The purity of ricin was determined as a percentage of the total protein in each lane, using the average density analysis tool.
Coomassie blue staining
Manufactured by GE Healthcare
Sourced in Sweden
Coomassie blue staining is a laboratory technique used to detect and visualize proteins in gel electrophoresis. It is a colorimetric method that binds to proteins and produces a blue color, allowing for the quantification and identification of proteins in a sample.
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Lab products found in correlation
Gs 800 calibrated densitometer, by Bio-Rad (2 mentions)
Polyacrylamide gradient 4 14, by GE Healthcare (2 mentions)
Coomassie blue staining, by Bio-Rad (2 mentions)
Quantity one software, by Bio-Rad (2 mentions)
Novex 4 20 tris glycine miniprotein gels, by Thermo Fisher Scientific (2 mentions)
Imperial protein stain, by GE Healthcare (2 mentions)
4 protocols using coomassie blue staining
1
Quantitative Analysis of Ricin Purity
2
Protein Separation and Visualization
3
Protein Separation and Visualization
4
Quantification of Ricin Purity by SDS-PAGE
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The purity of the ricin preparations was assessed by SDS PAGE using coomassie blue staining (GE Healthcare). Samples (1μL at 5 or 10 mg/mL) were mixed with a 1:1 volume of sample buffer Tris-HCl (125 mM) pH to 6.8, SDS (10%, w/v), glycerol (50%, v/v).
Samples were heated at 95°C for 5 min prior to loading 1 μL onto the gel (polyacrylamide gradient (4-14%); GE Healthcare). Proteins were separated for 40 min before coomassie blue staining for visualization, using manufacturer instructions (Calibrated Densitometer GS-800; Bio-Rad, Hemel Hempstead, UK). Densitometric scans were analyzed using Quantity One software (Bio-Rad). The value of the ricin band was determined as a percentage of the total protein in each lane, using the average density analysis tool.
Samples were heated at 95°C for 5 min prior to loading 1 μL onto the gel (polyacrylamide gradient (4-14%); GE Healthcare). Proteins were separated for 40 min before coomassie blue staining for visualization, using manufacturer instructions (Calibrated Densitometer GS-800; Bio-Rad, Hemel Hempstead, UK). Densitometric scans were analyzed using Quantity One software (Bio-Rad). The value of the ricin band was determined as a percentage of the total protein in each lane, using the average density analysis tool.
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