rna-isolation-purification-tissue-mouse-trachea

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 3D-Gene® Mouse miRNA Oligo chip (ver.21) to perform Microarray Gene expression arrays - Mouse liver tissue Cyanine-3-CTP

Get tips on using GeneChip® HT 3' IVT PLUS Reagent Kit to perform Microarray RNA amplification & Labeling - Mouse brain tissue Biotin

Get tips on using AllPrep Bacterial DNA/RNA/Protein Kit (50) to perform DNA isolation / purification Bacteria - Gram negative Massilia sp

Get tips on using RNeasy PowerBiofilm Kit (50) to perform RNA isolation / purification Biofilm

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 RNeasy Mini Kit to perform RNA isolation / purification Cells - immortalized Raji

Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - immortalized Raji

Get tips on using Anti-LC3B antibody produced in rabbit to perform Autophagy assay cell type - Human tracheobronchial epithelial cells (hTEC)

Get tips on using ISOGEN to perform RNA isolation / purification Cells - immortalized CHO