And NAO is the same that 10-Nonylacridine orange bromide?
Edited by cardosopedro, 14 November 2009 - 11:46 AM.
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#1
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Posted 14 November 2009 - 11:44 AM
I want to check if I have autophagy in drug-treated cells. I have nonyl acridine orange (NAO) available for the determination of mitochondrial. Recently I found that acridine orange can be used to detect autophagic vacuoles. Is there any difference between NAO and the acridine orange used for autopgahy detection? Or can I use NAO?
And NAO is the same that 10-Nonylacridine orange bromide?
And NAO is the same that 10-Nonylacridine orange bromide?
#2
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Posted 16 November 2009 - 06:30 PM
And NAO is the same that 10-Nonylacridine orange bromide?
Yes it is. This dye is usually used as a mitochodrial probe (not for autophagy)
Acridine orange is used in autophagy assays. It crosses into lysosomes (and other acidic compartments) and becomes protonated. The protonated dye stacks and stacked acridine orange emits in the red range. Acridine orange not in an acidic compartment will emit as green.
Yes it is. This dye is usually used as a mitochodrial probe (not for autophagy)
Acridine orange is used in autophagy assays. It crosses into lysosomes (and other acidic compartments) and becomes protonated. The protonated dye stacks and stacked acridine orange emits in the red range. Acridine orange not in an acidic compartment will emit as green.
#3
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Posted 17 November 2009 - 06:02 AM
Thanks.
So I won't be able to detect autophagy vacuoles with NAO, right?
So I won't be able to detect autophagy vacuoles with NAO, right?
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Posted 24 March 2010 - 02:14 PM
miBunny, on Nov 17 2009, 04:30 AM, said:
And NAO is the same that 10-Nonylacridine orange bromide?
Yes it is. This dye is usually used as a mitochodrial probe (not for autophagy)
Acridine orange is used in autophagy assays. It crosses into lysosomes (and other acidic compartments) and becomes protonated. The protonated dye stacks and stacked acridine orange emits in the red range. Acridine orange not in an acidic compartment will emit as green.
Yes it is. This dye is usually used as a mitochodrial probe (not for autophagy)
Acridine orange is used in autophagy assays. It crosses into lysosomes (and other acidic compartments) and becomes protonated. The protonated dye stacks and stacked acridine orange emits in the red range. Acridine orange not in an acidic compartment will emit as green.
Do you have idea on what concentration of AO is good for detecting autophagy in cells using FACS. In literature there is from 100 ng/ml to 100 mcg/ml, incubated for 15 min. Also, is it required to use compensation for detecting green and red signal from AOl? Because there is signal in all channels, from green to extreme red.
Is it possible to detect the autophagy proteins using FACS? Example: detecting LC3B, Beclin, ATG 5 or 12 or 7, using the primary antibodies (used for western blot), conjugated with fluoresence labelled secondary ab?
Thanks
