Blogs for February 2019

The LC3 A, B, C’s and 1, 2, 3’s

Tuesday, February 26, 2019 - 13:38
Atg5 expression in HeLa cells, ICC, immunocytochemistry, immunofluorescence

By Christina Towers, PhD

Autophagy is a catabolic process used to breakdown and recycle damaged proteins and organelles. It is a multistep process that, in its simplest form, consists of 4 steps: initiation, phagophore nucleation, elongation, and fusion – resulting in a double membrane structure called an autophagosome that, upon fusion with the acidic lysosome, facilitates the degradation of its contents. LC3B (and its family members) are incorporated into the elongating autophagophore and are necessary for efficient autophagosome...

Best way to quantitatively measure Autophagic Flux

Tuesday, February 19, 2019 - 10:36
LC3B expression in HeLa cells, ICC, immunocytochemistry, immunofluorescence

By Christina Towers, PhD

Autophagy is a stress-induced cellular recycling process that plays an important physiological role in many diseases. It is induced by a variety of stimuli, both intracellular and extracellular, and is a tightly regulated process with over 30 core autophagy proteins necessary for efficient autophagosome formation and fusion with lysosomes to facilitate degradation1. One of the most common ways to...

Wet or dry protein transfer: Which one to choose for your western blotting?

Tuesday, February 12, 2019 - 10:50
Novus Biologicals western blot resources page

By Jamshed Arslan, Pharm. D., PhD.

Western blotting involves three steps: 1) separating proteins by size through gel electrophoresis 2) transferring separated proteins to a solid support (typically, a PVDF membrane) and 3) using antibody to mark your protein(s) of interest in the mixture of proteins. The second step of moving negatively charged proteins from the gel to a blotting membrane (PVDF) can be performed in dry or wet conditions.

Dry/semi-dry transfers

Dry conditions are famous for easiness and speed. Reagents, power packs and wiring involved in dry transfers are less messy and therefore, potentially less hazardous. They allow blotting of several...

Animal Models to Study Autophagy

Tuesday, February 5, 2019 - 09:03
Autophagy Signaling Pathway, Novus Biologicals, macroautophagy induction, elongation, microautophagy, autophagosome maturation, autolysosome

By Christina Towers, PhD

What is autophagy?

Autophagy is the catabolic process that degrades cytoplasmic material via the lysosome. The process of macroautophagy was originally characterized in yeast, where the core autophagy-related proteins (ATGs) were identified. In this system, the multistep process of phagophore initiation and elongation as well as the conjugation machinery that mediate efficient autophagosome completion were determined and cloned to assess function. The mechanisms of starvation-...

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