siRNA / miRNA gene silencing Human WI-38

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.

When extracting nucleic acids from cell cultures, thorough homogenization of cells via vortexing in lysis buffer is very necessary. Choose the best RNA isolation method keeping in mind the downstream applications, generally, column-based isolations result in clean and concentrated RNA samples. Downstream applications like sequencing and cDNA synthesis require high-quality RNA, always treat the samples with DNases and check their integrity by running a gel.

Get tips on using QuikChange II XL Site-Directed Mutagenesis Kit, 10 Rxn to perform Site Directed Mutagenesis (SDM) Human - Deletion K562 c-Myb gene

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.

Get tips on using Nucleofector™ Kits for Human T Cells to perform DNA transfection Mammalian cells - Immortalized cell lines iPSC

Get tips on using Proteome Profiler™ Human Apoptosis Array Kit to perform Apoptosis assay cell type - Array of apoptotic proteins

Get tips on using Purified Mouse Anti-Human MSH-2 Clone G219-1129 (RUO) to perform Immunohistochemistry Human - MSH2

Get tips on using Monoclonal Mouse Anti-Human Progesterone Receptor (Concentrate) Clone PgR 1294 to perform Immunohistochemistry Human - PR

Get tips on using Monoclonal Mouse Anti-Human CA 125 (Dako Omnis) Clone M11 to perform Immunohistochemistry Human - CA125

Get tips on using Monoclonal Mouse Anti-Human CDX2 (Dako Omnis) Clone DAK-CDX2 to perform Immunohistochemistry Human - CDX2