NUN buffer
NUN buffer is a solution that makes it possible to purify proteins located in the nucleus of eukaryotic cells.[1] Although other procedures are available[2] 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.
References[edit]
- ^ Lavery, Daniel J.; Schibler, Ueli (1993). "Circadian transcription of the cholesterol 7 alpha hydroxylase gene may involve the liver-enriched bZIP protein DBP". Genes & Development. 7 (10): 1871–84. doi:10.1101/gad.7.10.1871. PMID 8405996.
- ^ Gorski, K.; Carneiro, M.; Schibler, U. (1986). "Tissue-specific in vitro transcription from the mouse albumin promoter". Cell. 47 (5): 767–76. doi:10.1016/0092-8674(86)90519-2. PMID 3779841. S2CID 26448485.