Protein isolation Tissue - Human placental 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.

Start discussion

Found 1 discussion for this experiment

Discussion

1 year ago

1 year 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?

Share your thoughts or question with experts in your field by adding a discussion!

Found 2 matching solutions for this experiment

Protocol tips
- Proteins are extracted with a Qiagen AllPrep DNA/RNA/Protein mini kit. Tissues are homogenized following the manufacturer’s instructions with two modifications: (i) addition of protease inhibitors to the lysis buffer (RLT) and (ii) the re-suspension of protein pellets in Modified Reswell Solution.
Protocol tips
- Placental tissue is homogenized using lysis reagent according to 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