Ettan daltsix ettan dalttwelve dige gel and dige buffer kit typhoon fla 9000 decyder 2d software v7. Final progress report for research projects funded by. Ingel digestion was performed after washing the gel plugs twice with 50 mm ammonium bicarbonate, then 50% vv. Genetixs gelpix robot combines the functionality of a gel imager and spot picker. To use the ettan spot picker or ettan spot handling workstation, two adhesive. Plant immunity induced by cosoga elicitor is a cumulative. Itr v acknowledgement ay 201011 pdf acknowledgement form 15, andor return of fringe benefits in form no. Briefly, ief electrophoresis was performed using immobilized ph gradient strips 18 cm, ph 310 nonlinear. Directions inautomated sample preparation of proteins. Compatibility of deep purple total protein stain with. Files are available under licenses specified on their description page. Sigmaaldrich offers a number of ge ettan spot picker products.
Spot picking with the ettan spot picker requires that gels are precast on backing e. For the second dige set of experiments, the proteins of interest were excised from the. The montage ingel digestzp kit, suitable for manual and. The easy way to treat glass plates with bind silane so that.
Ge health care ettan spot handling workstation robotic. The 2 mm diameter gel plugs were destained twice in 100 mmoll ammonium bicarbonate nh 4hco 3, 50% vv acetonitrile acn for 10 min. Peptides were chromatographically resolved online using a c18 col. Suitably diluted samples of the bladder homogenates mediated changes noted above but also in other respects 3032. User manual decyder differential analysis software, version 5. Data file 28402117 aa image analysis ettan dige imager. Comparison of pandemrix and arepanrix, two ph1n1 as03. Typhoon fla 9000 quick start guide baylor university. Ettan spot picker, ettan digester, ettan 2d system proteomics solution 1 system, symbiot i system spot picker proteomelab pa 800 and du 800 systems biomek fx and biomek2000 liquid handler biacore 3000 surface plasmon resonance ms system proteomeworks system massprep, clonis cellular lcm. Sigma sdependent antioxidant defense protects stationary.
Digestion and spotting onto maldi targets were performed in. Protein separation by 2de, image acquisition and analysis twodimensional gel electrophoresis 2de was done as previously described 14. For safe use of ettan dige system related products described in this user manual, see corresponding manuals. Ge healthcare ettan spot picker federal equipment company. Aerogel, an introduction the gel is used to get aerogel. The steam locomotive was in its early stages of development and the raincoat had yet to be sewn. The machine is kind of large it will occupy much of the space on a workbench and comes with two different sized spot picking heads.
To scan a gel with fluorescently labelled proteins. The procise 492 protein sequencer performs automated edman sequencing. Proteomic comparisons of opaque and transparent variants. Syprostained gel using the automated ettan spot picker gehealthcare into 96well plates and ingel digestion was carried out on the massprep digestion robot waters, manchester, uk. The protein picklist is transferred to ettan spot picker, which then automatically picks the protein spots according to the list. Research open access a novel function for vimentin. To spot pick from a cydye dige fluor minimal dye labelled gel, the gel must be poststained with sypro ruby to enable visualization of the. Place the strip in the ettan multiphor ii such that the gel side is up. Automated edman protein sequence analysis automated edman chemistry is performed on an applied biosystems 492 procise protein sequencer. The gel spots were washed, and digested ingel with modified porcine trypsin protease trypsin gold. Ingel digestion was performed as described by chan et al. Proteins were excised from the gels using the ettan spot picker ge healthcare, little chalfont, uk. Please change the country on your profile in order to switch to another country store. Imagequant las 4010 4000 4000 mini quick start guide.
Small rna mediated repression of subtilisin production in. Differential ingel electrophoresis in a highthroughput environment. Data file 28402117 aa image analysis the ettan dige imager fig 1 is a scanning ccd camera. Your comments optional i agree to the terms and conditions. Mavrc microscopy and imaging core, page 4 ettan spot picker the ettan spot picker is an excellent tool for picking spots from two dimensional protein gels 2d gels. Explore aerogel with free download of seminar report and ppt in pdf and doc format. Mavrc microscopy and imaging core denver, colorado. Alternatively, the integrated ettan spot handling workstation performs spot. The bigger head allows for coarse picking of spots and it picks up gel pieces with a diameter of approximately 1. The incidence of malignant melanoma mm was occurring at a faster rate than for most neoplasm. Deep purple, ecl, ecl advance, ettan, hybond, hyperfilm, imagequant, imagemaster. Data can also be exported in an xml format for either multigel analysis in decyder differential. Sample insoluble pellet 1 insoluble pellet 2 40 mm tris supernatant 1 8m urea, 4% chaps, 2mm tbp, 0. Itr i for assessment year, having the following particulars.
Extracted peptides were subsequently dried to reduce volume and remove organic solvent. Two different types of backing may be used, either. Allows quick and accurate protein spot picking from nonbacked 2d gels, increasing the versatility of ettan spot picker. All structured data from the file and property namespaces is available under the creative commons cc0 license. The ettan spot picker from amersham biosciences is an excellent tool for picking spots from two dimensional protein gels 2d gels. Serum depletion induces changes in protein expression in. Compatibility with ettan spot picker flat field calibration compatible with image analysis software, such as imagemaster, imagequant, or decyder software for. Ettan spot picker nonbacked gel kit cytiva formerly ge. The spots of interest were picked up by ettan spot picker ge healthcare based on the ingel analysis and spot picking design by decyder software. The procise is a complete, automated chemistry and analytical system for amino acid sequence analysis of protein and peptide samples in picomole to nanomole amounts. Ettan spot picker performs accurate and reliable picking of gel. This page was last edited on 29 august 2011, at 14. Proteomic analysis reveals the mechanisms of m ycena. Ettan spot picker instrument control software, running under windows nt operating system on a pc.
Ettan dige system user manual 18117317 edition ab 9 1 introduction 1. Here the gas is used in place of the liquid part of the gel. Gel spots were excised as 2mm diameter plugs and delivered into 96well microplates. It is a powerful tool for separating complex mixtures of proteins by charge and size and for scanning and analyzing the resulting second. The spots were excised using an ettan spot picker ge healthcare. Ettan spot picker user manual 18114761 ettan digester user manual 18116731 ettan spotter user manual 41 ettan spot handling workstation user manual 18115355.
Typhoon fla 7000 quick start guide vanderbilt university. Ettan snus its swedish for number one imagine what life was like in the year of 1822. Classical versus preserved speech variant alessiocortelazzo, 1,2 robertoguerranti, 1 claudiodefelice, 3 cinziasignorini, 4 silvialeoncini, 2,4 alessandrapecorelli, 2,4 claudialandi, 5 lucabini, 5 barbaramontomoli, 2 claudiasticozzi, 6 luciaciccoli, 4 giuseppevalacchi, 6,7. Working on the same principle as picking from backed gels, it uses reference markers to ensure high picking accuracy.
Spots were ingel digested with trypsin, separated on an ultimate 3000 nanohplc dionex, vienna, austria and analyzed by nanospray mass spectrometry using an ltq orbitrap xl, thermo finnigan thermo electron, vienna, austria as described in ref. The ettan comes with two different sized spot picking heads. Trypsinized peptides from two preparative gels were pooled and analyzed with nanolcesimsms maxis 4g uhrtof mass spectrometer bruker coupled with a 2dlc dionex ultimate 3000 thermo scienti. Used ge healthcare ettan spot handling workstation robotic. Automating 2d gel electrophoresis the scientist magazine. A spot picking list was generated from decyder software and exported to ettan spot picker. Pdf detection and mass spectrometry indentification of. Ettan dige system users manual university of alberta.