Storage of B-gal staining buffer
mattman at leland.stanford.edu
Fri Jul 28 18:13:06 EST 1995
In case anyone's curious...
(I'm posting this because SUMIT CHANDA told me to...)
Since some people store their B-gal yellow staining solution cold, some
store it in the dark, some store it on the bench in the light, and some
make it up fresh each time, I decided to compare them all. I made up
a batch of the stuff and split it into several bottles. One was kept
dark at 4 degrees, one was on the bench in a brown glass bottle, one
was on the bench covered with foil, and one was on the bench in a clear
glass bottle. Two months later, I stained LacZ expressing C2C12
myoblasts with all of the above solutions and also with a freshly
made batch. The results, amusingly, were that all gave identical
staining except for one; the FRESH batch gave inferior staining.
Which shows two things: (1) there can be batch to batch variability
depending on the alignment of the planets, and more importantly,
(2) storage in the light at room temperature is just fine over 2 months.
I think I'll try it again in about half a year...
(Staining solution was 5 mM K ferricyanide, 5 mM K ferrocyanide,
2 mM magnesium chloride in PBS pH 7.4. X-gal was added fresh to
1 mg/ml. Cells were fixed in 4% formaldehyde/0.25% glutaraldehyde
for 4 min before staining.)
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