ソーラーバイオクーマシー (ブラッドフォード) プロテインアッセイキット 50T

Cat No.: PC0015

サイズ: 50T

ストレージ: Seal preservation after opening, this kit from the date of order is valid for nine months.

説明

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.

プロトコル

• 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 回, 4 回, 8 times dilution), and add 20ul sample to 96 良い. 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 分.
  6. Determination of absorbance.
  7. According to the standard curve calculate the sample protein

• UV-Vis分光光度計

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 分

関連製品:

PC0001 Protein standard solution (5mg/ml BSA) PC0021 BCA Reagent

PC0030 Lowry Protein Assay Kit PC0020 BCAプロテイン Assay Kit R0010 RIPA buffer(high)

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

Related Paper:

• Zhongyuan Li,Xiumei Li,Tianhui Liu,他. The critical roles of exposed surface residues for the thermostability and halotolerance of a novel GH11 xylanase from the metagenomic library of saline-alkaline soil. International Journal of Biological Macromolecu Les. JuLy 2019;133:316-323. (IF784)
• Qinlu Zhang,Qian Liu,Menghan Du,他. Cetuximab and Doxorubicin loaded dextran-coated Fe3O4 magnetic nanoparticles as novel targeted nanocarriers for non-small cell lung cancer. Journal of Magnetism and Magnetic Materials. June 2018. (IF046)