"A question of Life or Death" - differentiating between healthy and apoptotic cells
Dead or Alive on the ZE5™ - Using flow cytometry to measure apoptosis
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The progression down the orderly process of autophagy can be an early indication of poor cell health due to starvation or growth factor suppression, ER stress, infection or immune signals.
We offer a range of antibodies and kits to detect and measure markers of autophagy. These include reagents to detect the lysosome, the autophagosome and the most studied macroautophagy pathway.
For more information on autophagy and a more comprehensive list of autophagy antibodies including the ATG proteins responsible for regulation of the autophagy machinery visit our dedicated autophagy page.
Figure 1. Staining of staurosporine-induced THP-1 cells using the Magic Red Cathepsin B kit
Autophagy Probe Red is a cell-permeant aliphatic molecule that fluoresces brightly when inserted into the lipid membranes of autophagosomes and autolysosomes. Fusion of the autophagosome with a lysosome to form an autolysosome is crucial to allow enzymes such as cathepsins to hydrolyze cargo for recycling back to the cytoplasm.
A quick and easy method is to determine Cathepsin B, K and L levels in lysosomes of both adherent and suspension cells is to use our Cathepsin Green and Magic Red kits™
Figure 2. Western blotting analysis of LC3 expression on Atg5 knockout and wild type cells using Rabbit anti Human MAP1LC3A/B (AHP2167). Image courtesy of Dr. R. Kopito, Stanford University.
We have antibodies to detect common markers of autophagy. These include Beclin-1 which is one of over 30 autophagy proteins involved in the regulation of the pre-autophagosomal structure. Another reliable method of monitoring autophagy is to use the mammalian homologue of ATG8, MAPILC3A/B. These ubiquitin-like proteins are found in autophagosomes. Autophagy can be measured by changes in cellular localization and detection of two bands by Western blot due to the presence of cytosolic LC-I and PE bound LC-II. The macroautophagy pathway can be studied using antibodies against lysosome-associated membrane proteins Lamp 1 & 2 (CD107a and b).
Autophagy is only one aspect of cell health. We have antibodies, reagents and kits to detect viability, proliferation and apoptosis to have confidence in your cell health.
Magic Red™ is a trademark of Immunochemistry Technologies, LLC.