BCA Working Reagent Recipe:
BCA reagent A: 50 mg/mL of bicinchoninic acid in 2% sodium carbonate solution.
BCA reagent B: 4% cupric sulfate solution in 0.4 N sodium hydroxide.
Mix 50 parts of BCA reagent A with 1 part of BCA reagent B. Mix well and incubate at room temperature for 30 minutes before use.
In duplicate, dispense 50µL of each sample or diluted protein standard to be treated into a 2.0mL microcentrifuge tube.
2. Add 500µL of Compat-Able Protein Assay Preparation Reagent 1 to each microcentrifuge tube. Mix each tube and allow the tubes to stand at room temperature for at least five minutes.
Compat-Able Protein Assay Preparation Reagent 1
What is the standard solution for Bradford assay?
Bradford Assay Materials: BSA standard solution (0.1 µg/µl) •
Bradford solution o
Dissolve 100 mg Coomassie Brilliant Blue G-250 in 50 ml 95% ethanol. Add 100 ml of 85% phosphoric acid while stirring continuously. When the dye has dissolved, dilute to 1 l in H2O.
Allow the vial and reconstitution buffer to equilibrate to room temperature. Briefly centrifuge or tap down the vial to ensure that all lyophilisate is collected at the bottom of the vial. Add the amount of buffer required to achieve the concentration recommended on the product insert or Certificate of Analysis.
How to reconstitute reagents?
The process of reconstitution involves adding a specified volume of distilled water to lyophilised QC material. The water should completely dissolve the lyophilised contents, giving a liquid solution, which is ready for analysis. Reconstitution is a straightforward process, but requires a high level of precision
What is the meaning of Lyophilisate?
Definitions of lyophilisation. a method of drying food or blood plasma or pharmaceuticals or tissue without destroying their physical structure; material is frozen and then warmed in a vacuum so that the ice sublimes. synonyms: freeze-drying, lyophilization.
What is the process of buffer preparation?
Common preparation methods include: 1) dripping an acid (or alkali) into an aqueous solution of a salt while measuring the pH with a pH meter and 2) making an aqueous solution of acid with the same concentration as the salt and mixing while measuring the pH with a pH meter.
Albumin Standard Ampules, 2mg/mL
Dilute the contents of one Albumin Standard (BSA) ampule into several clean vials, preferably using the same diluent as the sample(s). Each 1 ml ampule of 2.0 mg/ml Albumin Standard is sufficient to prepare a set of diluted standards for either working range suggested in Table 1.