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Sections were developed in staining solution (detergent solution plus 5 mM K3Fe CN 6, 5 mM K4Fe CN 6, 5 mM ethylene glycol tetraacetic acid (EGTA), 1 mg/mL X-gal (5-bromo-4-chloro-3-indolyl-β D-galactoside, Invitrogen, Carlsbad, CA) at 37 °C for several hours or overnight.
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Glutamate containing lipid-detergent vesicles can be prepared using crude soybean phospholipids (18), a glutamate containing solution, the detergent β-octylglucoside, and, for detergent removal, gel filtration spin columns ((19); Andersen SSL, unpublished data).
Ott first took a rat kidney and stripped out its functional cells using a solution of detergent.
Next, the proteins and fats that make up the cellular and nuclear partitions are broken down and removed with a solution of detergent.
This step helps to lyse the cells using lysing solution with detergent.
Samples were centrifuged and digested in a solution of detergent (sodium dodecylsulfate) and proteinase K, for removal of proteins bound to the DNA.
Sample equipment was cleaned in a 1% solution of detergent and hot water between sample collection to prevent cross contamination.
A 1% (w/v) solution of detergent was prepared, and the substrates were first cleaned by sonicating them in this solution for 10 minutes using an ultrasonic bath.
The platinum mesh basket electrode was cleaned with an aqueous solution containing detergent (Decon 90) for 20 min in an ultrasonic cleaning bath (180 W, 40 kHz).
Results were calculated as: % Lysis = Experimental-Effector Spontaneous-Target Spontaneous X 100 Target Maximum-Target Spontaneouswhere target maximum represents target cells plus Promega lysis solution containing detergent.
Mix up a solution of detergent.
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