Protein concentrations were determined by the densitometry of bands on SDS-PAGE gels stained with Coomassie Blue R-250 using carbonic anhydrase as a standard and a ChemiDoc MP imaging system (Bio-Rad Laboratories). Fluorescent proteins were detected by direct in‐gel fluorescence imaging using the same ChemiDoc imaging system. Alexa532 and Alexa647 concentrations were determined using ε531 = 81,000 cm−1 M−1 and ε650 = 270,000 cm−1 M−1, respectively. Western blot bands were visualized by chemiluminescence using the Clarity Max Western blotting kit (Bio-Rad Laboratories) and the ChemiDoc imaging system. IMV concentrations were determined as the A280 in 2% SDS [1 (link)]. All error bars are standard deviations.
Clarity max western blotting kit
Manufactured by Bio-Rad
The Clarity Max Western blotting kit is a product from Bio-Rad designed for the detection and quantification of proteins in western blot analysis. It provides the necessary reagents and materials for performing the western blotting technique.
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3 protocols using clarity max western blotting kit
1
Protein Quantification and Visualization
2
Protein Concentration Determination Methods
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Concentrations of purified proteins were determined by the densitometry of bands on Coomassie Blue R-250 stained SDS-PAGE gels using carbonic anhydrase as a standard and a ChemiDoc MP imaging system (Bio-Rad Laboratories). Fluorescent proteins were detected by direct in‐gel fluorescence imaging using the same ChemiDoc imaging system. Alexa647 and mNeon concentrations were determined using ε650 = 270,000 cm−1 M−1 and ε506 = 116,000 cm−1 M−142 (link), respectively. Western blot bands were visualized by chemiluminescence using the Clarity Max Western blotting kit (Bio-Rad Laboratories, #1705062) and the ChemiDoc imaging system. IMV concentrations were determined as the A280 in 2% SDS7 (link).
3
Protein Quantification Using SDS-PAGE
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Protein concentrations were determined by the densitometry of bands on SDS-PAGE gels stained with Coomassie Blue R-250 using carbonic anhydrase as a standard and a ChemiDoc MP imaging system (Bio-Rad Laboratories). Fluorescent proteins were detected by direct in-gel fluorescence imaging using the same ChemiDoc imaging system. Alexa532 and Alexa647 concentrations were determined using e 531 = 81,000 cm -1 M -1 and e 650 = 270,000 cm -1 M -1 , respectively. Western blot bands were visualized by chemiluminescence using the Clarity Max Western blotting kit (Bio-Rad Laboratories) and the ChemiDoc imaging system. IMV concentrations were determined as the A 280 in 2% SDS (1) . All error bars are standard deviations.
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