Home About us Contact
|
Home About us Contact | ||
|
|
||
Purified Fraction (purified + fraction)
Selected AbstractsCharacteristics of monoclonal antibodies against Piscirickettsia salmonisJOURNAL OF FISH DISEASES, Issue 4 2001A Jamett A panel of 28 monoclonal antibodies against Piscirickettsia salmonis was produced using a purified fraction of the bacterium. To determine their specificity to the pathogen, the antibodies were assayed by ELISA and indirect immunofluorescence microscopy. Six monoclonal antibodies were selected based on their strong reaction against P. salmonis and absence of cross-reactivity with other common fish pathogens. Western blot analysis showed that the antibodies reacted to several antigens of P. salmonis. Immunofluorescence assays revealed that these antibodies reacted with the same specificity to different isolates of P. salmonis obtained from the south of Chile. This panel of monoclonal antibodies represents an important tool to develop simple, rapid, sensitive and highly specific methods for the detection of the pathogen and diagnosis of the disease. [source] Optimization of enzymatic extraction of ferulic acid from wheat bran, using response surface methodology, and characterization of the resulting fractionsJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 10 2009Hélčne Barberousse Abstract BACKGROUND: The agro-industries generate thousands of tons of by-products, such as bran or pulps, each year. They are, at best, used for cattle feeding. Through biocracking, this biomass may constitute a renewable source for various molecules of interest for the industry. For instance, ferulic acid, a compound showing antioxidant ability, is found in abundance in cereal bran. Its release depends mainly on the breaking of its ester linkage to other constitutive elements of the cell wall, such as arabinoxylans. Response surface methodology was used to evaluate the effects of ferulic acid esterase (FAE) and xylanase activities, as well as incubation time and temperature, on ferulic acid extraction yield from wheat bran. Under optimized conditions, the composition of the hydrolysate and of residual bran were compared to native bran. RESULTS: Experiments carried out under the predicted optimal conditions (FAE amount, 27 U g,1; xylanase amount, 304 U g,1; incubation time, 2 h; and temperature, 65 °C) led to an extraction yield of 52.8%, agreeing with the expected value (51.0%). The crude ferulic acid fraction was purified with Amberlite XAD16, leading to a final concentration of 125 µg mL,1 of ferulic acid in ethanol. The antioxidant capacity of this purified fraction was evaluated by the DPPH· scavenging method: it exhibited better efficiency (EC50 = 10.6 µmol L,1 in ferulic acid) than the ferulic acid standard (EC50 = 13.7 µmol L,1). CONCLUSION: These results confirm the potential of wheat bran valorization in the field of natural antioxidant extraction, possibly viable in an industrial scheme. Copyright © 2009 Society of Chemical Industry [source] Identification of oleuropein oligomers in olive pulp and pomaceJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 10 2006Susana M Cardoso Abstract Analysis of a purified fraction from acetone extracts of olive pulp and pomace by electrospray ionisation tandem mass spectrometry (ESI-MSn) showed the presence of oleuropein oligomers, whose occurrence has not been reported previously in the literature. The main ionic species (m/z 1613) in the ESI-MS spectrum was an oleuropein trimer containing three linkages through the hydroxytyrosol backbone. In both samples, oleuropein dimers (m/z 1075), trimers comprising two hydroxytyrosol linkages (m/z 1615), tetramers (m/z 2153) and pentamers (m/z 2691) were also detected by MS. The occurrence of oleuropein oligomers was also observed by nuclear magnetic resonance (NMR). 13C, 13C distortionless enhancement by polarisation transfer DEPT 90, 13C DEPT 135, gHSQC (heteronuclear single quantum coherence) and gHMBC (heteronuclear multiple bond coherence) spectra showed all carbon and proton resonances of oleuropein with the exception of the low-mobility and asymmetric signals of the aromatic rings. Since mature olives were used in this study, it is possible that the disappearance of oleuropein that has been described to occur with the olive fruit maturation, could be associated with the formation of phenolic oligomers together with lower-molecular-weight compounds resulting from its degradation. Copyright © 2006 Society of Chemical Industry [source] Purification and identification of a transcription factor, USF-2, binding to E-box element in the promoter of human telomerase reverse transcriptase (hTERT)PROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 2 2010Shoulei Jiang Abstract Controversy remains about the identity of the transcription factor(s) (TFs), which bind to the two E-box elements (CACGTG, proximal and distal) of the human telomerase (hTERT) gene promoter, the essential elements in the regulation of telomerase. Here, systematic oligonucleotide trapping supplemented with 2-DE and proteomic methods was used to identify E-box binding TFs. Although insufficient purity was obtained from the proximal E-box element trapping, further fractionation provided by 2-DE and specific identification from Southwestern blotting analysis allow us to clearly identify an E-box binding TF. The protein spot was cut from 2-DE and in-gel digested with trypsin for LC-nanospray ESI-MS/MS analysis. This identified upstream stimulatory factor 2 (USF2). Western blotting analysis with specific antibodies clearly shows USF2 present in the purified fraction and USF2 antibody supershifts the specific DNA-binding complex on non-denaturing gels. Furthermore, a novel method was developed in which the specific DNA-TF complex was separated on a non-denaturing gel, the band was cut and applied to SDS-PAGE for a second dimension. Western blots of this second gel also confirmed the presence of USF2. [source] Phenolic compounds in the brown seaweed Ascophyllum nodosum: distribution and radical-scavenging activitiesPHYTOCHEMICAL ANALYSIS, Issue 5 2010Laetitia Audibert Abstract Introduction , Phenolic compounds are metabolites exhibited at high levels in Phaeophyceae. Although several studies have been conducted on total phenol contents, no one to our knowledge has dealt with the contents of phenolic compounds and antioxidant activities on purified fractions. Objective , The purpose of this study was the extraction and purification of phenolic compounds from the brown seaweed Ascophylllum nodosum, to determine both their distribution and their radical-scavenging activities, and to obtain a sufficiently purified oligophenolic fraction to perform an RP-HPLC analysis on molecules with a molecular weight (MW) < 2,kDa. Methodology , Phenolic compounds were separated and purified by liquid,liquid extraction, tangential ultrafiltration and dialysis. Then, the contents of both phenolic compounds and radical-scavenging activities were measured by the Folin,Ciocalteu reagent, and DPPH and ABTS assays. NMR analysis was performed to validate the process. RP-HPLC with a C18 column was performed on the oligophenolic fraction, using a novel method developed in this study. Results , Seven fractions were obtained as a function of polarity and molecular weight. Among them, the fraction containing phenolic compounds with a MW , 50,kDa appeared to be the most active, correlated with the content of phenolic compounds. Conclusion , This work constitutes a step forward in the separation and purification of bioactive phlorotannins and represents a prerequisite for further investigations into their structural characterisation and distribution in A. nodosum. [source] | ||||||||||