Xtractor buffer composition
In this latter case, the chitin fraction obtained also had a significant residual protein content. The possibility of using an enzymatic assisted extraction of proteins was also explored, obtaining a maximum nitrogen solubilisation in the best case (with Bacillus licheniformis protease) of about 60%. To avoid protein damage in alkali media, a stepwise protein extraction adopting milder conditions was also explored based on Osborne fractionation method, allowing the recovery of > 85% of BSF high purity and high quality proteins, and the obtainment of chitin-enriched fraction as well. The best separation was obtained by alkali extraction of proteins (96% of protein recovered) albeit with loss in their integrity as indicated by the measurement of the degree of hydrolysis with the o-phthaldialdehyde method. The lipid fraction was easily recovered by organic solvents, while the most challenging issue was the separation of protein from chitin. High buffer capacity of the composition in a wide pH range, 2.5-10 units pH, is provided by a complex of buffer systems: in the acidic range of pH, 2. BSF prepupae biomass contained, expressed on dry weight, 32% proteins, 37% lipids, 19% minerals, 9% chitin.
![xtractor buffer composition xtractor buffer composition](https://www.takarabio.com/images/800-Protein/CC-TechFigs/xTractor-Buffer-extraction-schematic-9126-iT.gif)
The clarified lysate (12 L) was incubated with PBS, 10 nM VC Sia, or 10 nM VC Sia and 100 nM BT4244 for 1.5 h at 37 C. First, exact proximate composition, total amino acids, fatty acids profile, and N-acetylglucosamine content of the prepupae samples were determined. COSMC KO K562s (2 × 10 6 cells) were spun down, washed twice with PBS, and lysed in 200 L of xTractor buffer supplemented with 2 L of 100× EDTA-free protease inhibitor and 10 U of DNase I. The present investigation was undertaken to evaluate the feasibility of applying different extraction protocols, either chemical extractions or enzymatic assisted extraction, to recover pure fat, protein and chitin fractions. TALON xTractor Buffer: No preparation necessary except. protein sample is diluted with Loading Buffer and loaded onto the column, and. Before preparing other buffer compositions, please consult AppendixA to evaluate resin. Black soldier fly (BSF, Hermetia illucens) constitutes an economic way to convert residual biomasses into a valuable source of biomolecules, such as proteins, lipids and chitin. Overview of the Phosphopeptide Enrichment protocol.