Issue 047 • 3/08/2011 Hot, Fast Protocol for FFPE-FASP™ Looking to save time in the lab? Use Protein Discovery’s FFPE-FASP Protein Digestion Kit to de-paraffinize, un-crosslink, solubilize, acetylate, and deplete your biobank samples for digestion. Our unique, hot extraction buffer method means incubation takes just 30 minutes — half the time of published protocols! Download [...]
Mass Spectrometry Sample Prep Product News and Updates
Recent News and Information Related to Protein Discovery’s extraction, solubilization, quantification, fractionation and digestion products for mass spectrometry analysis of proteins.
Protein Discovery has just added six new institutions to the company’s growing list of customer organizations: Iwate University Okinawa Institute of Science & Technology Kyoto University Graduate school of Pharmaceutical Science Tokyo University Laboratory for Systems Biology and Medicine Children’s Hospital Seattle University of Texas Health Science Center at San Antonio Welcome! We appreciate your [...]
Plant Physiol Biochem . 2011 Feb 18; Schacht T, Unger C, Pich A, Wydra K Polygalacturonases (PGs) of wild-type and non-virulent phenotype conversion mutant (PC) strains of Ralstonia solanacearum were compared by investigating their activities and their inhibition by polygalacturonase-inhibiting proteins (PGIPs) from tomato stems. In cultures of wild-type strain ToUdk2, slimy (s), retarded slimy (rs) and non-slimy (ns) colonies appeared. The conversion of the 's' into the 'rs' colony form coincided with the beginning of PG production. PG activity of the PC strain increased about 5 h earlier (at 6 hpi), and was up to 35 times higher in media supplemented with two different tomato stem extracts or polygalacturonic acid, compared to the wild-type at 6 hpi, and generally 4-8 times higher across test media and time. By hydrophobic interaction chromatography (HIC), fluorophor-assisted carbohydrate-polyacrylamid-gel electrophoresis (FACE-PAGE) and mass spectrometry analyses, endo-PG PehA, exo-PGs PehB and PehC were identified. PGs of the PC mutant consisted mainly of endo-PG. The increased PG production after supplementing the medium with tomato cell wall extract was reflected by a higher activity of exo-PGs for both strains. Total PGs (endo-PG and exo-PGs) activities were inhibited by PGIPs of tomato stem extracts. PGIP activity was concentration dependent, constitutively present, and not related to resistance nor susceptibility of tomato recombinant inbred lines to R. solanacearum. The proteinaceous character of the inhibiting component was inferred from ammonium sulphate precipitation. For the first time a plant PGIP activity against a bacterial pathogen is reported. Observations support that endo- and exo-PG synthesis is governed by a sensitive regulatory network, which, in interaction with PGIP and cell wall degradation products, leads to generation or avoidance of elicitor-active oligomers, and, thus, may contribute to the development of the compatible or incompatible interaction.
Looking to save time in the lab? Use Protein Discovery’s FFPE-FASP Protein Digestion Kit to de-paraffinize, un-crosslink, deplete SDS and other contaminants, and digest formalin-fixed paraffin-embedded FFPE tissue samples. Our unique, hot extraction buffer method means incubation takes just 30 minutes — half the time of published protocols! Download the FFPE-FASP Protein Digestion Kit protocol (pdf).
As promised, on February 1, Protein Discovery did indeed select the winner of the iPad giveaway event announced in the January edition of their “Sample Prep” newsletter. In an e-mail to this webmaster, veep Andrea Mravca wrote: “At dinner tonight, we put all the names in a box and had our waiter pick a winner [...]
Mol Biosyst . 2010 Apr; 6(4): 677-9 Abe R, Caaveiro JM, Kudou M, Tsumoto K N-Acylamino acids are a new family of versatile biological surfactants capable of extracting integral membrane proteins of various topologies from the biological membrane, in many instances surpassing the efficiency of commercial detergents.
Rapid and efficient protein digestion using trypsin-coated magnetic nanoparticles under pressure cycles. Proteomics. 2011 Jan;11(2):309-18 Authors: Lee B, Lopez-Ferrer D, Kim BC, Na HB, Park YI, Weitz KK, Warner MG, Hyeon T, Lee SW, Smith RD, Kim J Trypsin-coated magnetic nanoparticles (EC-TR/NPs), prepared via a simple multilayer random crosslinking of the trypsin molecules onto magnetic nanoparticles, were highly stable and could be easily captured using a magnet after the digestion was complete.
Related: Improving Protein Digestion for Shotgun Proteomics, PPS Silent Surfactant Flaubert Mbeunkui, Michael B. Goshe To evaluate the implementation of various denaturants and their efficacy in bottom-up membrane proteomic methods using liquid chromatography-mass spectrometry (LC-MS) analysis, microsomes isolated from tomato roots were treated with mass spectrometry-compatible surfactants (RapiGest SF Surfactant from Waters Corporation and PPS Silent Surfactant from [...]