rna-isolation-purification-cells-primary-rat-brain-microvascular-endothelial-cells

The challenge in isolating RNA from S. aureus cells is the disruption of the cell wall. A lot of protocols employ enzymatic digestion (pretreatment) which may affect gene expression patterns of certain genes. Therefore physical disruption using beads is considered to be the better alternative.

Get tips on using TRI Reagent® Sigma to perform RNA isolation / purification Cells - primary human cardiac fibroblasts

Get tips on using Gentra Puregene Cell Kit to perform DNA isolation / purification Cells - Primary cells Human primary keratinocytes

Isolating RNA from tissues and paraffin-embedded tissue samples can be challenging due to cross-linking of biomolecules and fragmented nucleic acids. The best solution is to slice the tissues into smaller pieces and make a homogenate solution (using tissue homogenizer or grinding liquid nitrogen frozen samples) in presence of RNAse inhibitors. The homogenization process should be carried out on dry ice to maintain the integrity of RNA.

Isolating RNA from tissues and paraffin-embedded tissue samples can be challenging due to cross-linking of biomolecules and fragmented nucleic acids. The best solution is to slice the tissues into smaller pieces and make a homogenate solution (using tissue homogenizer or grinding liquid nitrogen frozen samples) in presence of RNAse inhibitors. The homogenization process should be carried out on dry ice to maintain the integrity of RNA.

Get tips on using Pierce™ Cell Surface Protein Isolation Kit to perform Protein isolation Mammalian cells - Human aortic endothelial cells

Get tips on using RNeasy Micro Kit to perform RNA isolation / purification Cells - primary mouse dorsal root ganglion neurons

Get tips on using RNeasy Plus Mini Kit to perform RNA isolation / purification Cells - primary human peripheral blood monocytes

Get tips on using RNeasy Plus Mini Kit to perform RNA isolation / purification Cells - primary human osteoblasts - rheumatoid arthritis

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.