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Position: Discussion forums > Bioscience biotechniques discussion forums > Molecular biology troubleshooting forum
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Concentration of Triton X-100
Posted by: genefan (IP Hidden, New member, 1)
Date: June 6, 2005 10:15PM
Today I lysed pancrease islet cells using lysis buffer with 0.1% Triton-X100. But I have found most protocols use 1%. The lysis buffer contains 50mM Tris.Cl, pH7.5, 150mM NaCl. Could anyone tell me if it is OK? Thank you.
Re: Concentration of Triton X-100
Posted by: femmeauburn (IP Hidden, Advanced member, 115)
Date: June 7, 2005 12:47PM
the best way to tell if the lysis buffer was okay is to measure the amount of protein in the obtained samples. if the detergent concentration was too low poor lysis/homogenization will be evident by the lowered protein yield. incidentally, one of the lysis buffers I myself have used successfully did not contain Triton X100 or another detergent at all.
Re: Concentration of Triton X-100
Posted by: 5'-ATCG (IP Hidden, Unregistered user, )
Date: June 7, 2005 06:54PM
The 1% Triton-X100 is correct since islet cells are tightly aggregated, compact cell clusters.
0.1% tritonX-100 will not work either for protein or DNA/RNA extraction.
Re: Concentration of Triton X-100
Posted by: femmeauburn (IP Hidden, Advanced member, 115)
Date: June 8, 2005 11:57AM
the tissue type makes all the difference in chosing a lysis buffer. if your tissue type is tightly aggregated, compact cell clusters like 5'-ATCG indicates, then 0.1% will not work. I am not certain if there is any way to 'save' your samples at this point.
Re: Concentration of Triton X-100
Posted by: Rebekah Vaughn (IP Hidden, Unregistered user, )
Date: July 22, 2005 02:03PM
Anybody have any suggestions on a Triton X-100 alternative for a gram-positive bacteria lysis buffer?
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