Bioepitope^RBradford Protein Assay Kit

Introduction and product overview:

Bradford coomassie-binding, colorimetric method for total protein quantitation is one of dye binding methods.CoomassieBriliant Blue G-250 in the free stateshowsbrown, and itsmaximum light absorbance is 488nm; When CoomassieBriliant Blue G-250 combines with protein ,it turns brown into blue,and the maximum absorption is 595 nm.The absorbance at 595nm is proportional to the protein concentrations.Protein binding CoomassieBriliant BlueG250 reaches a balance in about 2 mins.The complex remains stable in 1 hour at room temperature. The method has superiority such as simple and convenient operation, sensitive reaction (4 times higher than Lowry Method), etc.

Biogot Bradford protein assay kit is developed based on Bradford coomassie-bindingmethod.This method has traits of simple and convenient operation，high sensitivity，accurate quantification and stable effect.

Kit contents:

BiogotCoomassieBradfordAssay Reagent：100ml，stored at 4℃。

Bovine Serum Albumin Standard: 100mg(Power), Stored at 4℃.(Recommend to make stock solution(5mg/ml) and aliquot before store at 4℃)

Test Tube Procedures:

A)      Standard Preparation:

Dilute stock solution from 5mg/ml to 0.5mg/ml. The diluent used should be the same as used for the protein samples.Label7 test tubes with A-G and prepare the standards as indicated below. The following dilutions are suitable for duplicate Standard assays.

B) Micro-plate Procedure (Sample to WR ratio = 1:20):

1.        Pipette each standard or unknown sample into the appropriate microplatewells.The sample would be less than 20µl,and add diluent up to 20 µl.

2.         Add 200μL of the BiogotCoomassie BradfordAssay Reagent to each well and mix with plate shaker for 30 seconds.

3.         Cover plate and incubate for 5 minutesat RT.

4.         Measure the absorbency at or near 595 nm on a plate reader.

5.         Use the standard curve to calculate the protein concentration of each unknown sample.

Notes:

1.        Make BiogotCoomassie BradfordAssay Reagentreturn to RT before use, and mix well.

2.        Please wear the lab coat and gloves to operate.

3.        Makesure the absorbency at 595nm within the range of the standard curve.

4.        Good linear range for samples is from 50-2000μg/ml.

5.        Period of validity is 6 months.