Protein isolation Mammalian cells - Mouse Epididymal 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.

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
- Mice epididymal white adipose tissue samples are homogenized in T-PER™ Tissue Protein Extraction Reagent with a protease inhibitor cocktail and a phosphatase inhibitor. Centrifuge the sample at 10,000 × g for 5 minutes to pellet cell/tissue debris.
Protocol tips
- Tissues are homogenized with the PRO-PREPTM protein extraction solution and incubated for 10 min on ice. The lysates are centrifuged at 11,000× g for 20 min at 4 °C.
RIPA Buffer

Sigma-Aldrich

Protocol tips
- Fat tissues are homogenized in 1 mL of RIPA buffer using a homogenizer.
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!

Outsource your experiment

Fill out your contact details and receive price quotes in your Inbox

  Outsource experiment
Become shareholder Discussions About us Contact Privacy Terms