Solarbio Coomassie (Bradford) Protein Assay Kit 50T

Cat No.: PC0015

Size: 50T

Skladištenje: Seal preservation after opening, this kit from the date of order is valid for nine months.

Opis

Coomassie G-250 dye is a colorimetric reagent used for the detection and quantitation of total protein. In an acidic medium Coomassie dye binds protein causing an immediate shift in absorption maximum from 465nm to 595nm with a concomitant color change from green to blue. In a certain concentration range, the absorbance value A595 measured is in direct proportion to the protein concentration.

Protocol

• MicroplateReader
  1. Completely dissolve BSA standard, dilute 10ul to 250ul, the final concentration of 0.2mg/ml. The dilution buffer was dependent on the measured protein sample. For the sake of simplicity, suggested to use 0.9%NaCl or PBS.
  2. 5×G250mix well before 1ml 5×G250 diluted with 4ml ddH2O. 1×G250 solution can save one week at 4℃.
  3. The standard according to 0, 2, 4, 6, 8, 12, 16, and 20ul respectively added to 96well plates, and PBS dilution to complement.
  4. Dilute the sample(prepare a few gradients, such as 2 times, 4 times, 8 times dilution), and add 20ul sample to 96 well. To avoid errors, sample points need set after the standard line of 1/2.
  5. Add 200ul diluted 1×G250 to each well, room temperature for 3-5 mins.
  6. Determination of absorbance.
  7. According to the standard curve calculate the sample protein

• UV-VisSpectrophotometer

1. 8*10ml centrifuge tube(or more) and labeled with1~8.
2. Dilute 100ul BSA with 2.4ml PBS to 2mg/ml.
3. 5×G250 mix well before use. 1ml 5×G250 diluted with 4ml ddH2O. 1×G250 solution can save one week at4℃.
4. Add reagents according to the following table (each well 5ml, the extra is used to clean the cuvette)

5. Measure OD value after 3 minutes of reaction. To ensure the accuracy of the experiment, measure the OD value every 2 minuta

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PR1600 Prestained Protein Marker(14.4kD-97.4kD) R0050 Nuclear Protein Extraction Kit

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