Autophagy assay cell type - B35

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

LC3B antibody

GeneTex

Upstream tips	Protocol tips	Downstream tips
	- For Immunoblot dilute Ab at overnight at 4°C in 1:500 dilution. - For IF dilute Primary Ab dilution-1:200 and incubate for 1h

Protocol tips
- For Immunoblot dilute Ab at overnight at 4°C in 1:500 dilution. - For IF dilute Primary Ab dilution-1:200 and incubate for 1h

Manufacturer protocol Publication protocol 1 Relevant paper

Anti-LAMP2 antibody [H4B4]

Abcam

Upstream tips	Protocol tips	Downstream tips
	- Primary Ab dilution-1:100 and incubate fro 1h

Protocol tips
- Primary Ab dilution-1:100 and incubate fro 1h

Manufacturer protocol Publication protocol 1 Relevant paper

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