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Team:SUSTC-Shenzhen/Notebook/A-B Toxin/Bradford
From 2014.igem.org
Bradford
a method to meter the concentration of protein
2014.9.2, 9:00~21:00
Material
Tiangen Bradford G-250 staining buffer, Tiangen 1mg/ml BSA, sample
Protocol
| add\number | 1 | 2 | 3 | 4 | 5 | 6 |
|---|---|---|---|---|---|---|
| BSA(1mg/ml) | 0 ul | 10 ul | 20 ul | 30 ul | 40 ul | 50 ul |
| ddH2O | 50 ul | 40 ul | 30 ul | 20 ul | 10 ul | 0 ul |
| stain | 950 ul | 950 ul | 950 ul | 950 ul | 950 ul | 950 ul |
2.incubate for about 0.5 hrs
3.use the spectrometer to get the data
4.compare the sample with the principle data
Select the 1st elution protein as the sampling sample
TEG: for 50uL, A=0.198-- c=2.66ug/L
GD5: for 50uL, A=0.129-- c=1.01ug/L
Evaluation
The concentration of protein is too low for us to us, especially after the recovery, the concentration will be less than 1ug/L. (we must have at least 10ug/L’s protein, or we can’t get enough useful protein)
Method
1. use the protein we made lately. 2. try to use the precious protein
