Protein isolation Tissue - Lamb muscle tissue

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.

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4 years ago

4 years ago by Jesualdo Hernández Spain

When should I be harvesting the tissue before using it for the most optimal results?

I am planning to do HIF nuclear extraction using mouse brain tissue. How fast should I be harvesting the tissue before using it for the most optimal results?

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Protocol tips
Total proteins of five muscle tissues were extracted by using a Total Protein Extraction Kit (Sangon Biotech, Shanghai, China), according to the manufacturer’s protocol, and then the muscle proteins were preserved at −80 °C in a freezer.
Protocol tips
Protein was extracted from snap-frozen liver and skeletal muscle (100 mg) into 2 ml of CelLytic MT reagent following the CelLytic MT mammalian tissues lysis procedure
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