5x bradford reagent preparation
Recipe Dissolve 50mg of Coomassie Blue G in 50ml of methanol Add ml of 85% . Bradford's Reagent Recipe Caution: Always add acid slowly into water and do not add water into acid. Preparation of reference solutions. Reference Protein. [. Concentration. The calibration curve should be created new for each series of tests. (e.g. BSA). When the dye comes in contact with protein, the first electron is donated to charged groups on the protein. The Bradford reagent is an acidified solution of Coomassie G; the dye is thus primarily protonated and red. The basis for the assay is that in order for the Coomassie dye to bind stably to protein, it needs to be doubly protonated. (Do not shake the bottle to mix the solution!). To perform the assay in microtiter plates dilute the 5x Bradford Reagent ,5 (2 parts by volume plus 5,5 parts by volume H 2 O bidest.). Please mix the reagent gently by inverting the bottle serveral times. Remove the amount of reagent needed and equili-brate it to room temperature before use. Recipe Dissolve 50mg of Coomassie Blue G in 50ml of methanol Add ml of 85% H 3 PO 4 to the solution from step 1 Add the solution from step 2 into ml of H 2 O and mix Filter to remove and precipitates Add an additional ml of H 2 O. Bradford's Reagent Recipe Caution: Always add acid slowly into water and do not add water into acid. 5. Incubate the mixture at room temperature for 5 minutes. After adding 25 μL of 5X Bradford Reagent to each microwell, gently pipetting or tapping microwell plate. 6. Measure the . 4. Recipe · Dissolve 50mg of Coomassie Blue G in 50ml of methanol · Add ml of 85% H3PO4 to the solution from step 1 · Add the solution from step 2 into ml of.