Team:USTC CHINA/Notebook/Protocols/Concentrating proteins
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<h1>Concentrating proteins</h1> | <h1>Concentrating proteins</h1> | ||
- | <p>Performed with Sangon Biotech(Shanghai) | + | <p>Performed with Sangon Biotech(Shanghai)</br> |
- | Protocol: | + | Protocol:</br> |
- | + | 1.Prepare a 1.5ml clean centrifuge tube and add 200ul sample protein solution to the tube.</br> | |
- | + | 2.Add 50ul precipitaton solution A and reverse the tube vertically for 10 seconds to make it mixed up evenly. </br> | |
- | + | 3.Put the tube on ice or the substratum of fridge and incubate it for an hour. </br> | |
- | + | 4.Centrifuge the tube for 15 minutes with 15000rpm, temperature 4℃. </br> | |
- | + | 5.Remove the supernatant and the liquid on the tube wall and tube bottom, and remain the sediment. </br> | |
- | + | 6.Add 600ul washing liquor and reverse the tube vertically for 10 seconds. </br> | |
- | + | 7.Put the tube on the substratum of fridge for 10 minutes and centrifuge the tube for 15 minutes with 12000rpm, temperature 4℃ to get the sediment.</br> | |
- | + | 8.Pour away the supernatant and dry out the sediment in the draught cupboard for 30 minutes or unwater the sediment with freeze drier. </br> | |
- | + | 9.Add 20ul solution C and reverse the tube vertically for 10 seconds; if the solution turns yellow, add 1-5ul buffer solution to make it blue </br> | |
- | + | 10.Boiling the solution for 5 minutes and centrifuge it for 5 minutes with 12000rpm at room temperature.</br> | |
- | + | 12.Sample the supernatant and carry out SDS-PAGE electrophoresis and analyze the molecule weight.</br> | |
Revision as of 14:51, 25 September 2013