|
Materials |
Procedure |
Microassay |
Materials
required/used for protein assay.
Spectrophotometer (595 nm)
Cuvettes with 1 cm path length matched to lab spectrophotometer
13 x 100 mm test tubes
Test tube rack
Vortex mixer
Whatman #1 filter and funnel
Graduated cylinders
Pipets 100ul and 5ml
1. Prepare dye reagent by diluting 1 part Dye Reagent Concentrate with 4 parts distilled, deionized water. Filter through Whatman #1 filter to remove particulates.
2. Prepare three to five dilutions of a protein standard, which is representative of the protein solution to be tested. The linear range of the assay for BSA is 0.2 to 0.9 mg/ml.
3. Pipet 100 ul of each standard and sample solution into a clean, dry test tube. Protein solutions are normally assayed in duplicate or triplicate.
4. Add 5.0 ml of diluted dye reagent to each tube and vortex.
5. Incubate at room temperature for at least 5 minutes. Absorbance will increase over time; samples should incubate at room temperature for no more than 1 hour.
6. Measure absorbance at 595 nm.
1. Prepare three to five dilutions of a protein standard which is representative of the protein solution to be tested. The linear range of the assay for BSA is 1.2 to 10.0 ug/ml.
2. Pipet 800 ul of each standard and sample solution into a clean, dry test tube. Protein solutions are normally assayed in duplicate or triplicate.
3. Add 200 ul of dye reagent concentrate to each tube and vortex.
4. Incubate at room temperature for a least 5 minutes. Absorbance will increase over time; samples should incubate at room temperature for no more than 1 hour.
5. Measure absorbance at 595 nm.
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