What is an autophagy assay?

Table of Contents

Autophagy Assay Kit ab139484 measures autophagic vacuoles and monitors autophagic flux in live cells using a dye that selectively labels autophagic vacuoles. It can be analyzed using flow cytometry, fluorescent microscopy, or using a microplate reader.

Does autophagy result in cell death?

Although experimentally induced excessive autophagy can cause cell death in non-mammalian organisms and in cultured mammalian cells, we are not aware of any model system that convincingly shows that physiological (or even pathological) cell loss in mammalian cells in vivo is executed by autophagy.

Can autophagy be selective?

Depending on the cargo, autophagy can be selective or nonselective. In nonselective autophagy, bulk portions of the cytoplasm are sequestered by the phagophore for degradation to maintain the nutrient levels in cells. This type of autophagy is frequently observed upon low nutrient levels during starvation.

How is autophagy activity measured?

Autophagy can be monitored by two different approaches: (1) direct observation of autophagy-related structures and their fate; and (2) quantification of autophagy-/lysosome-dependent degradation of proteins and organelles.

What is an example of autophagy?

Autophagy has roles in various cellular functions. One particular example is in yeasts, where the nutrient starvation induces a high level of autophagy. This allows unneeded proteins to be degraded and the amino acids recycled for the synthesis of proteins that are essential for survival.

Does autophagy induce apoptosis?

Still yet, there are cases where autophagy seemingly induces cell death irrespective of apoptosis. A number of molecules can affect both autophagy and apoptosis, suggesting that they may act as molecular switches of these two cellular processes in response to cell damage signaling.

What is selective autophagy?

In the past three decades, selective autophagy has been characterized as a highly regulated and specific degradation pathway for removal of unwanted cytosolic components and damaged and/or superfluous organelles.

Is autophagy the same as apoptosis?

Apoptosis and autophagy are important molecular processes that maintain organismal and cellular homeostasis, respectively. While apoptosis fulfills its role through dismantling damaged or unwanted cells, autophagy maintains cellular homeostasis through recycling selective intracellular organelles and molecules.

How is autophagy flux measured?

Autophagosome flux can be quantified experimentally by completely blocking one of the steps in the pathway and measuring the initial rate of accumulation in the substrate for the inhibited step, such as, for example, the rate of autophagosome accumulation upon complete inhibition of the lysosomal fusion step.

Is autophagy necessary for autophagic cell death?

Recently, RNAi silencing for autophagy-essential genes has played a major role in molecularly establishing autophagy as necessary for autophagic cell death (table 2). This approach provides the molecular specificity that chemical reagents lack.

How can autophagy be assessed?

A wide array of methods can be used to assess autophagy. Morphological hallmarks of autophagy can be observed by transmission electron microscopy (EM) and confocal microscopy.

What is the role of autophagy in the detection of lysosomes?

Since autophagy is a constitutive cellular degradation process, the use of an autophagy detection reagent will prevent the lysosomal degradation of LC3, allowing its quantification by flow cytometry.

What are autophagy-related genes (ATGs)?

Autophagy-related genes (ATGs) regulate the autophagy and also control the crosstalk with autophagy-associated cell death and apoptosis in some condition. Various methods have been used to detect the marker genes and the proteins involved in these processes.