Protein isolation Mammalian cells - Mouse_Brown fat

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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Found 2 matching solutions for this experiment

Upstream tips
- Tissues were lysed in the RIPA buffer with protease inhibitor and protein phosphatase inhibitor.
Protocol tips
- Membrane protein extraction was followed by instructions of Mem-PER Plus Membrane Protein Extraction Kit according to manufacturer's protocol.
TRI Reagent®

Sigma-Aldrich

Protocol tips
- Total protein is isolated from Brown fat tissues using TRI Reagent following the manufacturer’s instructions.
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