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Get tips on using TRI Reagent® MRC to perform RNA isolation / purification Tissue - Rat Heart

Get tips on using TRI Reagent® MRC to perform RNA isolation / purification Tissue - rat heart tissue

Get tips on using pET28a-HsLARP6 to perform Protein Expression Prokaryotic cells - E. coli LARP6

Get tips on using X-VIVOTM 10 Serum-free Hematopoietic Cell Medium to perform Mammalian cell culture media NK-92

Get tips on using X-VIVOTM 10 Serum-free Hematopoietic Cell Medium to perform 3D Cell Culture Media Mouse thymic organoids

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.

ROS has a very short half-lives in biological environment as they are influenced by exposure to ambient oxygen. As it is highly reactive and hard to measure care should be taken to ensure the stability of the sample during isolation, preparation, storage, and analysis.