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Biotin picolyl azide reacts with alkyne to form 1,2,3-triazole by 1,3-dipolar Huisgen cycloaddition through the use of a much lower copper (I) concentration without sacrificing reaction efficiency.
Biotin picolyl azide reacts with alkyne to form 1,2,3-triazole by 1,3-dipolar Huisgen cycloaddition through the use of a much lower copper (I) concentration without sacrificing reaction efficiency.
Even though AccuOrange™ buffer does contain SDS, which is required for the dye to bind proteins, the assay is very sensitive to small changes in SDS concentration, and also cannot tolerate non-ionic detergents that form mixed micelles with SDS, like Triton®. Therefore we don’t recommend using the kit for cell lysates or other samples with significant amounts of detergents.
Gels stained with One-Step Blue® can be dried just like gels stained with Coomassie. The stain will not interfere with the detection of radiolabeled proteins.
The AccuOrange™ assay is a fluorescent dye-based assay. The dye binds to proteins primarily through hydrophobic interactions. Proteins denature upon heating; the dye binds to the exposed hydrophobic pockets of the protein after cooling. The free AccuOrange™ dye is fluorogenic due to non-radioactive decay but becomes highly fluorescent due to the rigid conformation inside the pocket.
The AccuOrange™ assay more sensitive than traditional protein quantitation assays such as BCA, Bradford and Lowry, and shows superior linearity and reproducibility than the NanoOrange® protein quantitation assay (Thermo Fisher Sci.), but has low tolerance for detergents like SDS and Triton® X-100.