Protein isolation Mammalian cells - Human aortic endothelial cells

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.

Start discussion

No discussions found

Start your discussion

Share your thoughts or question with experts in your field

Start a discussion

Found 3 matching solutions for this experiment

Protocol tips
The frozen tissues were homogenized in CelLytic™ MT cell lysis reagent (Sigma, C3228) by Precellys 24 beads homogenizer (Bertin Technologies, Montigny-le-Bretonneux, France). The homogenates were centrifuged and the supernatants were used for western blot analysis.
Protocol tips
- M-PER containing Halt™ Protease and Phosphatase Inhibitor is added to the cell pellets and proteins are extracted by gentle shacking for 10 min followed by centrifugation at 13,000× g for 15 min.
Protocol tips
- Follow the manufacturer's protocol.
Can't find the product you've used to perform this experiment? It would be great if you can help us by Adding a product!
Become shareholder Discussions About us Contact Privacy Terms