PROTEIN AND LPS GELS
REFERENCE: Hitchcock and Brown. 1983. Journal
of Bacteriology, 154:269-277.
OBJECTIVE: This is a procedure analyzing LPS produced
by bacteria, by using a protein-free LPS preparation and after gel electrophoresis,
staining the gel using a LPS specific stain.
0.1 M Tris-HCl, pH 6.8
Grow the bacteria overnight on appropriate agar plates.
Harvest cells with A sterile loop and suspend in 0.9% saline
to an OD600 of 0.4 (equivalent to 200 Klett units using a blue
Remove 1.5ml of the suspension and centrifuge in a microfuge
for 1 1/2 minutes to pellet the cells.
Solubilize the pellets in 100 µl of lysing buffer.
Heat the lysates to 100ºC for 10 minutes.
For protein digestion of the samples, add 25 µg of
proteinase K, solubilize in 10 µl of the lysing buffer to each boiled
lysate. Incubate at 60ºC for 1 hour.
Run the gel as usual and stain with silver (see the silver
Updated 01 December 2000.