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An Epitope Tagging Vector for Gene Expression in Mammalian Cells
...ne Expression in Mammalian Cells Use tag-specific antibodies to study expressed genes Tanya Hosfield Quinn Lu Stratagene The pCMV-Tag1 vector is an epitope tagging vector designed for geneexpression in mammalian cells. A target gene inserted into the pCMV-Tag1 vectorcan be tagged with the FLAG epitope ...Antibodies for Studying NMDA Receptor Protein Expression and Synapse-Specific Immunolabeling
New antibodies for neuroscience research Danny Q. Hoang Mic...ms, Inc. Stratagene introduces affinity purified antibodies for NMDA receptorsubunits that are useful for studying expression of the NR2A, NR2B and NR2Csubunit ...Epitope-Tagging Vectors for Functional Analysis in Yeast
...thin a cell, eliminating the need to make specific antibodies to eachnew protein under study. 9 To test the FL... the yeast lysates to Western blot analysis using antibodies specific to the epitope tag. A 64-kDa band, corresponding to the expected size of th...The Best Enzyme for the Toughest PCR Challenges Improved with New Hot-Start Feature
...merase is formulated with a combination of antibodies that effectively neutralize DNA polymerase and 3'-5' (proofreading) exonuclease activities. Full enzyme activity is regained once the antibody is denatured during ramp-up to initial denaturation temperatures. Enhance...Reliable Assessment of Micronuclei for Genotoxicity Testing in Mice or Rats
... (i.e., the CD71-defined antigen), and fluorescent antibodies directed against these surface markers can differentially label immature erythrocytes. 9,10 When fixed blood cells are treated with ribonuclease and antitransferrin receptor antibody (FITC-conjugate), and then stained...Simple, Sensitive Isotyping of Mouse Monoclonal Antibodies
...d assay,using colloidal gold-conjugated anti-mouse antibodies for isotype determinationof 0.1 g/ml of antibody in 5 minutes. For decades, antibodies have been used as key reagents for biologicalresearch. The development of hybridoma technology, whic...Greater Amplification Specificity with New Hot Start PCR Enzyme
... DNA polymerases reversibly inactivated by antibodies regain activity upon denaturation of the antibody (1 minute at 80C 8 ) during ramp-up to initial denaturation temperatures. In comparison, commercially available preparations of modified Taq remain inactive until high...Yeast Protein Production System Features High Yields and One-Step Purification
...G peptide (DYKDDDDK) 5 at its N terminus; antibodies against the FLAG peptide are available from Stratagene. Establishing the Expression Strain The gene of interest is inserted into the multiple cloning site of the pESP-1vector and is fused in frame to the upstream GST gene. Successful clonin...Enzyme Immunoassay for Studying Intracellular Levels of cAMP
... with goat anti-rabbit IgG. Rabbit anti-cAMP antibodies are combined with the sample and cAMP-horseradish peroxidase conjugate (tracer) and applied to the microtiter well. Samples containing cAMP compete with a fixed quantity of tracer for the rabbit anti-cAMP-specific antibodies. ...hNT BlueNeurons Express b-Galactosidase at High Levels
... identified in transplantedtissue by staining with antibodies specific for human markers, an additionalmethod of identification would allow immunostaining for multiple markers. ThelacZ gene was chosen as a marker to be introduced into hNT neurons because it isvery stable and does not appear to interfere with ce...Sample Preparation for Microinjection
...the highest activity of the particular proteins or antibodies should be used. For peptide antibodies raised in rabbits, affinity purification is recommended. The concentration of the materi...Principles and applications of electrical cell fusion
... Producing monoclonal antibodies Traceable to G. Khler and C. Milstein [4], this method revolutionized the production of antibodies. Primary B lymphocytes, which produce a specific antibody, are fused with tumor cells. The numerous fu...... hGH is usually carried out using 125 I-labeled antibodies against the growth hormone. Several manufa...e in the form of ELISA kits, with anti-hGH antibodies bound to the surface of a microtiter plate. First, the hGH from the supernatant of the cultu...Principles and applications of electrical cell fusion
... Producing monoclonal antibodies Traceable to G. Khler and C. Milstein [4], this method revolutionized the production of antibodies. Primary B lymphocytes, which produce a specific antibody, are fused with tumor cells. The numerous fu...... hGH is usually carried out using 125 I-labeled antibodies against the growth hormone. Several manufa...e in the form of ELISA kits, with anti-hGH antibodies bound to the surface of a microtiter plate. First, the hGH from the supernatant of the cultu...HotMasterAn innovative Hot Start/Cold Stop technology for better PCR* results
...ket today rely on chemical modification or antibodies to inhibit Taq polymerase; thus, their action is l...ns as do the Hot Start DNA polymerases that employ antibodies to achieve inhibition. Testing has also shown that HotMaster Taq can be used in real...TransMessenger Transfection Reagent
...and stained using mouse monoclonal anti-MAP2 antibodies and Alexa Fluor 488-conjugated goat anti-mouse secondary antibody (a-MAP2) and Alexa Fluor 594-conjugated phalloidin, which binds to f-actin and serves here as an expression control. Data kindly provided by A.M. Krichevsky and K.S. ...Effectene Transfection Reagent provides efficient gene delivery to primary neuronal cell cultures
...uclei (blue) of cells not reacting with MAP2 antibodies are glial-cell nuclei. Note that the nuclei of transfected neurons are not fragmented, which is an indicator of good cell health. In control transfection experiments (i.e., when the plasmid lacking the eukaryotic promoter was used),...Efficient DNA transfection of primary CNS neurons using TransMessenger Transfection Reagent
...cording to standard procedures using primary antibodies directed against MAP2 (a neuron-specific mar...rkers were detected using monoclonal primary antibodies (anti-MAP2 antibodies for neurons, anti-GFAP antibodies for astrocytes, the major subtype of g...LiquiChip Kits for bead-based cytokine and kinase assays
...of biotinylated cytokine-specific monoclonal antibodies and StreptavidinPE (Figures 1 and 2). The specific... threonine residues in MBP is detected using antibodies specific for phosphoserine and phosphothreonine. These antibodies are in turn detected using a...Transfection Reagent Selector Kit Handbook
...eted by transfected cells. Specific 125 I-labeled antibodies against hGH are used and results are monitored in a scintillation counter. Currently, a sandwich-ELISA is also often used, which involves an antibody coupled ELISA plate. The hGH protein binds to the antibody on the plate, a d...TransMessenger Transfection Reagent
...and stained using mouse monoclonal anti-MAP2 antibodies and Alexa Fluor 488-conjugated goat anti-mouse secondary antibody (a-MAP2) and Alexa Fluor 594-conjugated phalloidin, which binds to f-actin and serves here as an expression control. Data kindly provided by A.M. Krichevsky and K.S. ......fter transfection using mouse monoclonal anti-MAP2 antibodies and Alexa Fluor 488-conjugated goat anti-mou...ells were stained using mouse monoclonal anti-MAP2 antibodies and Alexa Fluor 488-conjugated goat anti-mouse secondary antibody (MAP2) and Alexa Fluor...Tools for Proteomic Science II
...ibody every time aprotein is studied. Our anti-Tag antibodies simplify thedetection, characterization, and purification of proteins.They are also available in conjugated forms, and recognizecommonly used epitope sequences, such as theinfluenza hemagglutinin protein (HA), human c-myc protein,six histidine residu......antibody is applied to theslides. Typically, these antibodies consist of Fab fragmentsthat are linked to alkaline phosphatase, peroxidase,or another enzyme for colorimetric detection(Roche Applied Science). An optional signal amplificationstep (TSA system, Perkin Elmer Lifesciences) helpsto detect transcripts o...Detection of mRNAs on Cryosections of the Cardiovascular System Using DIG-Labeled RNA Probes
... Alkaline phosphatase conjugated antibodies The sections were incubated with anti-DIG antibo...). Fluorochrome-conjugated antibodies When the mRNA of interest is abundantly expressed,anti-DIG antibodies conjugated with FITC or oth...Quantification of Nucleosomes in Serum by the Cell Death Detection ELISAplus
...wich-enzyme-immunoassay principle:Monoclonal mouse antibodies directed against DNA(single-stranded [ss] and doub...SA, 0.5% Tween, 1mM EDTAin PBS) for two hours. The antibodies were bound to thehistone- and DNA-component of the nucleosomes, andthe immunocomplexes were fixed to...In Situ Cell Death Detection Kit
...chemical detection using anti-fluorescein-specific antibodies that are conjugated to POD or AP. Typical applications of the TUNEL kits are: Detection of individual apoptotic cells in frozen and formalin-fixed tissue sections in basic research and routine pathology Deter......TO16. Staining of cell surface proteinswith antibodies may introduceartifacts that should be eliminatedby...osslinking ofseveral receptor molecules bybivalent antibodies may lead toreceptor internalization, where thereceptor becomes internalizedtogether with the bound a...Quality control of antibodies using the 2100 bioanalyzer and the Protein 200 Plus assay
...apeutics. Inall cases the successful production of antibodies requires precise testingand quality control. The k...n be usedas an ideal tool to study the stabilityof antibodies in stress testswith excellent reproducibility.This application is commonly usedas a quality control ...... positivecontrols. The specificity of the selected antibodies was determinedby flow cytometry. Results and discussion AdnaGen developed a diagnosticsystem for early detection ofdisseminated cells in peripheralblood of testicular, breast andcolorectal carcinoma patients.This diagnosti......croscopy using one of Ambion's new primary antibodies matched to our gene specific siRNA control...ontrols Ambion provides mouse monoclonal antibodies for the detection of GAPDH, c-myc, and -actin by immunofluorescence. Each of these primary a...Inducing RNAi with siRNA Cocktails Generated by RNase III
... after 48 hours, using specific primary antibodies (anti-GAPDH from Ambion; anti-La from Transduction Laboratories; anti-c-FOS from Santa Cruz Biotech). A FITC-conjugated donkey anti-mouse IgG secondary antibody (Jackson Immuno Research) ...Drive Long-Term Silencing with CMV Vectors
... was performed using gene specific primary antibodies and a fluorescein conjugated secondary antibody and the resulting reduction in protein expression was examined by fluorescence microscopy. The cells were mounted with DAPI to stain nuclei (Blue). Images were digitally captured...Isolate It All: siRNA miRNA Total RNA Native Protein
...e performed with 15 g of total protein and antibodies specific for GAPDH or Ku p70 (Ambion). Here, GAPDH knockdown was triggered by electroporation of a GAPDH-specific siRNA into a primary human cell line (normal human umbilical vein endothel......e performed with 15 g of total protein and antibodies specific for GAPDH or Ku p70 proteins (Ambion). Complete Kits The mir Vana Isolation Kits include sufficient reagents to perform 40 total RNA isolations or 20 enrichm...Superior Gene Silencing Using Adenoviral Vectors
...ortions and analyzed by Western blot using antibodies to GFP and -actin (control). Allele-spe...Ad-C10i virus. Western analysis was performed with antibodies to Ataxin-3 and GAPDH (as a control), and the siRNAs produced by the viruses were ab...The let-7 microRNA Family Regulates RAS: Implications for Development and Oncogenesis
...ancer patients. Western blots were performed using antibodies specific for RAS (US Biological) or GAPDH (Ambion) (top). Northern blot analysis was performed using radiolabeled probes for let-7 and U6 snRNA (middle). NRAS and -actin mRNA, as well as 18S rRNA (for normalizati...... levels were analyzed by Western blot with antibodies specific for F ocal A dhesion K inase, FAK (PTK2), or P roline-rich t yrosine k inase 2, Pyk2 (PTK2B). back to top Ordering Information ...... levels were analyzed by Western blot with antibodies specific for F ocal A dhesion K inase, FAK (PTK2), or P roline-rich t yrosine k inase 2, Pyk2 (PTK2B). back to top Ordering Information ...Purification of Murine IgG1 Using UNOsphere S and CHT Ceramic Hydroxyapatite Chromatography, Rev A
... as the first step in the process, yielding antibodies with very high purity in a single step. Ho... et al., Affinity purification of novel bispecific antibodies recognising carcinoembryonic antigen and doxorubicin, J Chromatogr B Biomed Sci and ......tein (25C9) and non-neutralizing monoclonal antibodies specific for epitope D of the S-protein (44C11) an...blot and the ELISA reaction with monoclonal antibodies (S, N), we demonstrated that immunogenic and neutralizing epitopes were preserved on t......ion is currently based on the detection of antibodies to distinct EBV antigens. In the past few ... enzyme-linked immunosorbent assay for IgM and IgG antibodies against the EBV-encoded protein p23. The recombinant antigen seems to be a useful di...... confirmed by western blot with polyclonal antibodies to bovine βLG. Immunoblotting shows that ^...olles, P., Antibovine β-lactoglobulin antibodies react with a human lactoferrin fragment and bovine β-lactoglobulin present in human mil...... IgG 1 -HRP or 1:1,000 goat anti-mouse IgG 2a -HRP antibodies (Serotec Ltd) in order to detect the subcl... all assessed by ELISA for the presence of antibodies to HBsAg. At all time points there was a significantly higher antibody titer compared to uni...Inoculation of Viral RNA and cDNA to Potato and Tobacco Plants Using the Helios Gene Gun
...orbent assay (DAS-ELISA), using monoclonal antibodies and alkaline phosphatase-conjugated monoclonal antibodies to PVA (MAb 58/0; Rajamaki et al. 1998) obtained from Adgen, UK. Optimization of Bom......ed beads (50 l) coated with target capture antibodies were transferred to each well of the filte... incubation and washing, premixed detection antibodies (50 l, final concentration of 2 g/ml) were added to each well. The incubation was terminated......hospho- and total protein ELISAs. The same antibodies specific to each target were used in both ...hosphoprotein and total target assays. The antibodies used in the ELISAs were those provided in the vendor kit. Results and Discu...Feasibility of Multiplexing Bio-Plex Total Target and Phosphoprotein Assays
...lly, each type of bead is conjugated with distinct antibodies that recognize one of the target proteins.... phosphoprotein assay. Also, because the pair of antibodies used in the two assays differ, the signal from a total target assay may not be comparable ...Proteomics in Bladder Cancer Research: Protein Profiling of Bladder Squamous Cell Carcinomas, Rev C
..., the tumor does not stain with keratin 19 antibodies (Figure 4B), is BG3C8-antigen positive (Figure 4E) (Celis et al. 1996b), keratin 13 negative (Figure 4H), keratin 14 positive (Figure 4K), and psoriasin negative, with the exception of the pearls (Figure 4N). Similar analysis ...Real-Time Immuno-PCR on the iCycler iQ System, Rev A
.... (1992). It combines the molecular recognition of antibodies with the high DNA amplification capability of PCR. The procedure is similar to conventional enzyme-linked immunosorbent assays (ELISA) but allows for more sensitive detection. Instead of an enzyme, a DNA molecule is li...The Bio-Rad Silver Stain, Rev E
...34. King SM et al., Characterization of monoclonal antibodies against Chlamydomonas flagellar dyneins by...85) 43. Linko-Kettunen L et al., Monoclonal antibodies to Bacteroides fragilis lipopolysaccharide, J Clin Microbiol 20, 519524 (1984) 44. Li......he same blot with minimal cross-reactivity between antibodies or dyes. Introduction ...ed) laser with a 670BP30 filter for Cy5-conjugated antibodies and the 532-nm (green) laser with a 580BP30 filter for Cy3 conjugates. The PMT value was adjusted un...The use of CypHer5 for receptor internalization studies in both a range of GPCRs and a non-GPCR
...body 1. Commercially available antibodies may be supplied in buffer containing sodium azide....gonist-induced internalization of CypHer5 labelled antibodies is concentration dependent. A sigmoidal dose response curve was obtained for each agonist by non-lin...The use of CypHer5E and the IN Cell Analyzer 1000 for live-cell receptor internalization studies
...gonist-induced internalization of CypHer5E-labeled antibodies was observed for all G-protein sub classes of GPCR...ti-EGFR antibody 1. Commercially available antibodies may be supplied in buffer containing sodium azide. This can affect labeling efficiency and therefore......body enable the immobilization of rabbit secondary antibodies during the assay. The Donkey Anti-Rabbit PS SPA Im...y principle Binding radiolabeled ligands to antibodies immobilized on the surface of an SPA Imaging Bead brings the radiolabel into close proximity with sc...Detection and quantitation of low-abundant proteins with ECL Plex fluorescent Western blotting
... Hybond-LFP membrane, and then probed with primary antibodies and secondary CyDyeconjugated antibodies according to the manufacturer’s instructions. Please see the application note Multiplex protei......say Kit has been developed with proprietary kinase antibodies and corresponding substrates to assess functional activity of a broad array of serine-threonine and tyrosine kinases capable of phosphorylating different kinase substrates (Table 1). <P > <P > Assay design ......h </LI> <LI >Concentration of antibodies and duration of incubations have been optimized so as to preserve the quality of RNA and to yield specific staining having a high signal/noise ratio. </LI> <LI >Addition of SuperaseIn RNase Inhibitor to the antibody cocktail...Laser Capture Microdissection of Cells Labeled with Enhanced Green Fluorescent Protein
...gy, histological stains or labeling by fluorescent antibodies in a heterogeneous tissue. Specificity of the LCM ...y, histological stains, or labeling by fluorescent antibodies from a heterogeneous population of cells in a tissue section. In this Application Note, an adaptatio...Other ContentsappendectomyarmpitarmpitarachnodactylyappetiteappetiteappetiteappetiteappetitearrhythmiaarrhythmiaarrhythmiaarrhythmiaarrhythmiasarrhythmiasarrhythmiasarrhythmiasOther TagsLuteinLupusLupusLupus
(Date:7/23/2008)...livestock manure into a domestic renewable fuel so...to three per cent of North America,s entire consum... greenhouse gas emissions (GHGs), according to US ...e Institute of Physics, Environmental Research Le...rgy and Emissions Benefits of Converting Manure to...
(Date:7/23/2008)...udy of parasites living in three estuaries on the ...esearchers have determined that biomass of these p...ses by more than 20 times. , Their findings, whi...l implications, appear in this week,s issue of the...ving matter that exists in a given habitat. It is ...
(Date:7/23/2008)...at the National Institute of Standards and Technol...he formation of clumps of proteins in protein-base...arifies the conditions under which scientists can ...easuring the formation of protein aggregates, a ma...ol and safety in biologic drug manufacturing. , ...
(Date:7/23/2008)...ity of Washington study of top-selling laundry pro...d dozens of different chemicals. All six products ... toxic or hazardous under federal laws, but none o.... , "I first got interested in this topic because... public restrooms and the scent from laundry produ...
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(Date:7/25/2008)...nCotter Releases Results of Fourth Annual Physicia...ey, DETROIT, July 25 /PRNewswire/ -- Sullivan, Co... annual survey of physician on-call pay rates and,...eport. The survey report,with data from 132 healt...,on-call pay practices and rates paid for 33 physi...
(Date:7/25/2008)...Kesson Corporation Executive, Robert J. Reilly, Jo... -- Aethon Inc.,( http://www.aethon.com ), the lea...ply chain logistics, today announced that Robert J...sident -- Chief Financial Officer., Reilly, who w... for,McKesson Corporation,s Specialty Pharmacy Div...
(Date:7/25/2008)...AD, Calif., July 25 /PRNewswire/ -- AutoGenomics, ...ion statement on Form S-1 with the,Securities and ...al public,offering of shares of its common stock. ..., all of which are expected to be offered by,AutoG...g, have not yet been,determined., J.P. Morgan Sec...
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