Proteins for NMR analysis: More ¹⁵N labeled protein from bacterial cultures
Poor growth, low levels of expression and insufficient labeled protein are the most frequent challenges faced when using minimal medium to prepare proteins for structural characterization by NMR analysis.
EnPresso® B Defined Nitrogen-free is a pre-sterilized, chemically-defined growth system that increases the yield of ¹⁵N-labeled proteins.
Join other researchers who have shared their positive results:
• Very easy to prepare – just dissolve the tablets and add NH4Cl
• Since there is no need to monitor OD before induction, the entire process is simplified
• Quality of the proteins was confirmed by their ability to bind to their co-factors
• The increase in product-specific yield made purification much easier
• Yields were surprisingly high
• Identical degree of labeling and equivalent spectra
• This was a huge step forward. We were able to produce ¹⁵N-labeled cytokine fragments – something that was previously impossible using M9
• There was a high level of ¹⁵N incorporation and 2D ¹⁵N-1H HSQC NMR spectrum showed no significant difference when compared to the spectrum from the M9-derived protein
• We were extremely impressed with a 13-fold improvement compared to previous yields