Autophagy assay cell type - INS-1E

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 3 matching solutions for this experiment

BECN1 Antibody (H-300)

Santa Cruz Biotechnology

Upstream tips
- Lyse cells in RIPA lysis buffer containing a complete protease inhibitor mixture and a protein phosphatase inhibitor.
Protocol tips
- Dilute Primary Ab -1:1000

- Incubate antibody overnight at 4°C
Upstream tips
- Lyse cells in RIPA lysis buffer containing a complete protease inhibitor mixture and a protein phosphatase inhibitor
Protocol tips
- Dilute Primary Ab at -1:1000 dilution.
Protocol tips
- Dissolve the inhibitor in distilled water
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