Mammalian Lysis buffer for B-gal Assay

Mimi Liao m.liao at student.unsw.edu.au
Thu Aug 19 21:29:02 EST 1999


Hi all,

I am currently lysing the CHO-K1 cells with lysis buffer containg 50 mM
Tris-Cl (pH8.0), 150mM NaCl, and 1 % Trioton-100. And i'm measuring the
B-gal content using ONPG colormetric assay. when i added the Na2HCO3 to
stop the reaction, the solution became very cloudy. However, the solution
cleared up o/n.

My questions are;
1) Do we really need to add the stop solution ? Can we just measure the
absorbance straight away without stopping the reaction ?
2)Are there any recipe for mammalian lysis buffer which are compatible with
B-gal assay ?



Thanks very much in advance




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