Autophagy assay cell type - N27 dopaminergic cells

As autophagy is a multi-step process which includes not just the formation of autophagosomes, but most importantly, flux through the entire system, including the degradation upon fusion with lysosomes, which makes it quite challenging for detection. There are several methods for detection in mammalian cells, including immunoblotting analysis of LC3 and p62 and detection of autophagosome formation/maturation by fluorescence microscopy, Currently, there is no single “gold standard” for determining the autophagic activity that is applicable in every experimental context, hence it is recommended to go for the combined use of multiple methods to accurately assess the autophagic activity in any given biological setting.

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Found 2 matching solutions for this experiment

Anti-LC3B antibody produced in rabbit

Sigma-Aldrich

Upstream tips	Protocol tips	Downstream tips
	- For Western Blot, dilute Primary Ab at 1:5000 and incubate at 4 °C overnight. - For Immunofluorescence dilute primary Ab at 1:500 and incubate for 1h at RT

Protocol tips
- For Western Blot, dilute Primary Ab at 1:5000 and incubate at 4 °C overnight. - For Immunofluorescence dilute primary Ab at 1:500 and incubate for 1h at RT

Manufacturer protocol Publication protocol 1 Relevant paper

Purified Mouse Anti-Nucleoporin p62 Clone 53/Nucleoporin p62

BD Biosciences

Upstream tips	Protocol tips	Downstream tips
	- Dilute Ab at 1:500 - Incubate Ab for 1 h at room temperature

Protocol tips
- Dilute Ab at 1:500 - Incubate Ab for 1 h at room temperature

Manufacturer protocol Publication protocol 1 Relevant paper

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