Team:SUSTC-Shenzhen/Notebook/A-B Toxin/Bradford

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Team SUSTC-Shenzhen

Notebook

Element of an endeavor

Contents


Bradford

2014 --a method to meter the protein concentration

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

1.Add as table describes

2.incubate for about 0.5 hrs

3.use the spectrometer to get the data

4.compare the sample with the principle data pricinple curve 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

Maintained by the iGEM team SUSTC-Shenzhen.

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