Lesson plan
NUN buffer
NUN buffer is a solution that makes it possible to purify proteins located in the nucleus of eukaryotic cells. Although other procedures are available they result in loss of albumin D-box binding protein (DBP) which is unwanted if nuclear signal pathways are to be investigated. Therefore, a new extraction procedure was developed in 1993 to increase recovery of nonhistone proteins using a (NUN) solution containing 0.3 M NaCl, 1 M urea, and 1% nonionic detergent Nonidet P-40, which destabilize salt bridges, hydrogen bonds, and hydrophobic interactions, respectively; resulting in a disruption of interaction between proteins and DNA. By incubating nuclei in NUN buffer and centrifuging the solution, the supernatant will therefore contain nuclear proteins. NUN buffer contains: HEPES [pH 7.6], Urea, NaCl, DDT, PIC 1 & 2, 1.1% NP-40, Sodium orthovanadate, β-glycerol phosphate and water.
Article summary
NUN buffer is a solution that makes it possible to purify proteins located in the nucleus of eukaryotic cells. Although other procedures are available they result in loss of albumin D-box binding protein (DBP) which is unwanted if nuclear signal pathways are to be investigated. Therefore, a new extraction procedure was developed in 1993 to increase recovery of nonhistone proteins using a (NUN) solution containing 0.3 M NaCl, 1 M urea, and 1% nonionic detergent Nonidet P-40, which destabilize salt bridges, hydrogen bonds, and hydrophobic interactions, respectively; resulting in a disruption of interaction between proteins and DNA. By incubating nuclei in NUN buffer and centrifuging the solution, the supernatant will therefore contain nuclear proteins. NUN buffer contains: HEPES [pH 7.6], Urea, NaCl, DDT, PIC 1 & 2, 1.1% NP-40, Sodium orthovanadate, β-glycerol phosphate and water.
Key claims
No claims recorded.
Discussion questions
- Summarise the article in your own words. Which single claim would you verify first, and why?
- Which claim in this article is most likely to be revised as the sources update? How would you check its current status?
- If two sources disagreed about one of these facts, what would you need in order to trust one over the other?
Bibliography
No sources recorded.
Key claims and bibliography are generated from the same tracked claims and sources as the article page.