CRISPR Mouse Deletion RMA cells

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.

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.

Get tips on using IntestiCult™ Organoid Growth Medium (Mouse) to perform 3D Cell Culture Media Mouse gastric cancer organoids

Get tips on using IntestiCult™ Organoid Growth Medium (Mouse) to perform 3D Cell Culture Media Mouse small intestinal organoids

Get tips on using Mouse TNF alpha ELISA Kit (ab208348) to perform ELISA Mouse - TNF-alpha

Get tips on using Mouse TNF-alpha Quantikine ELISA Kit to perform ELISA Mouse - TNF-alpha

Get tips on using Mouse Lipocalin-2/NGAL DuoSet ELISA to perform ELISA Mouse - NGAL/LCN2

Get tips on using Mouse Dkk-1 Quantikine ELISA Kit to perform ELISA Mouse - Dkk-1

Get tips on using Mouse ICAM1 ELISA Kit (CD54) (ab100688) to perform ELISA Mouse - ICAM-1/CD54

Protein isolation is a technique that involves isolation and/ or purification of protein from cells or tissues via chromatography or electrophoresis. The major challenges in protein isolation include: 1. The concentration of proteins in cells is variable and tends to be small for some intracellular proteins. Unlike nucleic acids, proteins cannot be amplified. 2. Proteins are more unstable than nucleic acids. They are easily denatured under suboptimal temperature, pH or salt concentrations. 3. Finally, no generalized technique/protocol can be applied for protein isolation. Proteins may have different electrostatic (number of positively or negatively charged amino acids) or hydrophobic properties. Therefore, protein purification requires multiple steps depending on their charge (a negatively charged resin/column for positively charged proteins and vice-versa), dissolution (using detergents) and unlike in the case of DNA and RNA, instead of using salts, proteins should be isolated by isoelectric precipitation.