Autophagy assay cell type - RSC96

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-Beclin 1 antibody (ab62557)

Abcam

Upstream tips	Protocol tips	Downstream tips
- Seed cells at 2 × 105/mL	- Primary Ab dilution-1:100 at 4°C overnight

Upstream tips
- Seed cells at 2 × 105/mL
Protocol tips
- Primary Ab dilution-1:100 at 4°C overnight

Manufacturer protocol Publication protocol 1 Relevant paper

LC3B Antibody

Novus Biologicals

Upstream tips	Protocol tips	Downstream tips
	- Dilute Primary Ab dilution to-1:50. - Incubate Ab for overnight at 4°C

Protocol tips
- Dilute Primary Ab dilution to-1:50. - Incubate Ab for overnight at 4°C

Manufacturer protocol Publication protocol 1 Relevant paper

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