Cy dye labeling software

Cy3 and cy5 dyes attached to oligonucleotide terminus. Amersham cydye dige fluors minimal dyes for 2d dige. The word cyanin is from the english word cyan, which conventionally means a shade of bluegreen close to aqua and is derived from the greek. Cy dye fluorochromes are one of the popular choices for fluorescent labeling in applications such as fluorescence microscopy, flow cytometry and fluorescent immunoassays. Citation data is made available by participants in crossrefs citedby linking service. However, both alexa fluor dyes were significantly more resistant to photobleaching than were their cy dye. Differential in gel electrophoresis an overview sciencedirect topics. The reason for using spyro ruby, which stains all protein in the gel, is that the cy dye labeling is carried out such that the extent of incorporation will be cy dyemole protein. Selective labelling of cellsurface proteins using cydye dige fluor minimal dyes asa hagnermcwhirter, maria winkvist, stephanie bourin, rita marouga research and development, ge healthcare bio. Biotium offers a variety of stains for different organelles and cellular structures.

Cyanine2 minimal dye for protein labeling, an equivalent of cy2 nhs ester minimal dye. Cy2, cy3 and cy5 conjugates for permanent mounting. Labeling software is a vital tool for the modern industrial workplace. Cy dye synthesis difference gel electrophoresis dige is the latest in a series of major advances in the eld of twodimensional 2d gel electrophoresis, which was originally introduced by o farrel 1. It can also be used for the calculations of any cy3cy5labeled singlestranded dna. It can also be used to conjugate other proteins or peptides. Mixnstain cyanine dye antibody labeling kits biotium.

Labeling l11s87c with the cy5 maleimide dye mlabeling buffer 50 ml. Dimethylformamide dmf has to be ordered separately. Biosynthesis offer cy3 fluorescent dye oligonucleotide labeling. Labeling hazards helps keep your workers aware of dangers such as arc flash and reminds them to take proper precautions. Includes sufficient reagents for labeling 65 g of sirna. They share the same chromophore structure and spectral properties, differing principally in their linkage chemistry. Generally, twice the labeling density will be achieved if the duration of the labeling reaction is extended to 2 hours at. Biosynthesis offers cy5 fluorescent dye oligonucleotide labeling.

Cy 2 conjugates have maximum absorptionexcitation at 492 nm and fluoresce with a peak around 510 nm in the green region of the visible spectrum like fitc conjugates 520 nm, but they are more photostable and less sensitive to ph changes than fitc. It has an absorbance maximum of 646 nm and an emission maximum of 662 nm. In this movie, we present a protocol for intact cells using cydye dige fluor. Thus, alexa fluor dyelabeled dna probes do not aggregate or precipitate, even in highsalt hybridization solutions. Analysis software difference gel electrophoresis dige is a relatively straightforward application of differential labeling of protein samples using fluorescent dyes. Cy5 conjugation kit cy5 labeling kit ab188288 uses a simple and quick process for cy5 labeling conjugation of antibodies. Quasar 570 replaces cy3, quasar 670 replaces cy5 and quasar 705 replaces cy5. Since the cy dye has a mw of about 580 da, low mw proteins e. Selective labelling of cellsurface proteins using cydye dige. The anomalous peaks, previously observed for protein conjugates of the cy5 dye, are presumably due to the formation of dye aggregates.

Cydye dige fluor labelling kit for scarce samples ipmb. Scroll down or select an organelle below to learn more. Thiol monoreactive zwitterionic and pi balancing cy5 dye zcy5 dye. The minimal dyes are intended for general 2d application use where sufficient. Difference gel electrophoresis imaging and analysis.

The solid line connects average values and demonstrates the trend. Apexbio cy5 nhs ester dye for labeling aminecontaining. Learn about our antibody labeling kits and their advantages to conjugate an antibody to cy5 using this kit. Difference gel electrophoresis imaging and analysis of dige. Optimal fluorescence is found at 6 to 8 cy3 or cy3. More selective method of antibody labeling that reduces the possibility of the label interfering with subsequent antibodyantigen reactions. Cydye dige fluor minimal labeling kit ge healthcare, 25. This reagent is specially quantified each package contains specified amount of. We recommend a standard reaction for 1 hour at 37c. Alternative programs can also be applied according to the suppliers of gels and. Absorption spectra of protein conjugates prepared from these dyes showed prominent blueshifted shoulder peaks for conjugates of the cy dyes but only minor shoulder peaks for conjugates of the alexa fluor dyes. Later on when software performs some data analysis this results into accurate.

The dyes were a modification of cyanine dye indocyanine green icg which was used for angiography since 1970s, and they all contain two indolenine rings flanking polymethyne chain. Flexible for use in typical protein labeling protocols. Cydye dige fluor minimal dyes and saturation dyes are size and charge matched fluorescent dyes specifically designed for detecting protein abundance. So now in this technology the dige technology those cyanine dyes are used. Cy3, cyanine fluorescent dye oligonucleotide labeling. Quantitative comparison of longwavelength alexa fluor dyes. Label the intact cells with 600 pmol cydye dige fluor minimal dyes for 20. These labels can be incorporated at many positions in the fluorescent peptide or labeled peptide. Dyecpg and phosphoramidites are available for labeling oligonucleotides. Cydye dige fluor minimal dyes from ge healthcare biocompare. Usually, they are incorporated at the nterminus or at cyslys side chains. Amersham cydye dige fluors minimal dyes for 2d dige reagents for labeling protein with cy2, cy3 and cy5 dyes, before 2dimensional separation. Quasar 570, quasar 670 and quasar 705, with emission maxima at 570, 670 and 705 nm, respectively.

Cydye dige fluor labeling kit for scarce samples plus preparative gel labeling containing. So this method the dige method involves protein labeling and separation. This protocol is rapid, simple to use, and all three cydye dige fluor minimal dyes cy 2, cy 3 and cy 5 can be used to label cellsurface proteins. Cydye dige fluor minimal labeling kit ge healthcare, 258010. The quasar dyes may be used as direct replacements for the cy dyes and are anticipated to perform equivalently to their cy dye counterparts.

Amersham cydye dige fluor labeling kit for scarce samples. Ge healthcare amersham cy3, cy 5 maleimide monoreactive. Cy dyes are excellent alternatives to most other fluorescein dyes as they are brighter and offer greater photostability. Oct 22, 2012 selective labeling of cellsurface proteins using cydye dige fluor minimal dyes. Ge healthcare amersham cy3, cy 5 maleimide monoreactive dye. This reagent can be used to attach cy5 fluorophore to proteins and peptides containing cysteine residues, as well as to other thiolated molecules such as thiolcontaining oligonucleotides. Cydye dige fluors are available as minimal and saturation labeling dyes. Cy5 conjugation kit easy cy5 labeling ab188288 abcam. In addition to immunocytochemistry applications, cy3 dye is commonly used to label nucleic acids. Amersham cy 3, cy 5 maleimide monoreactive dye packs are packaged in premeasured amounts and foilsealed to ensure consistent labelings. Cyanine2 minimal dye for protein labeling, an equivalent of cy2. Labeling of the surface proteins in living escherichia coli and staphylococcus aureus cells.

For the 2d dige, an li wt sample labeled with cy5 and an li mutant sample labeled with cy3 were run on the same gel. As nhydroxysuccinimidyl ester dyes, the alexa fluor 555 dye was similar to the cy3 dye, and the alexa fluor 647 dye was similar to the cy5 dye with respect to absorption maxima, emission maxima, stokes shifts, and extinction coefficients. Cydye postlabelling reactive dye pack provides conveniently packaged, highly reactive cy3 and cy5 monofunctional nhs ester to label probes via the post labeling amino allyl route for use in microarray applications. A, b twodimensional gel fractionation of cy dye labeled proteins from e. It contains all reagents required for performing 10 separate labeling reactions of 1 mg of poi. The protein samples the one labeled with cy3 dye, the other labeled with cy5. In addition, fluorescently labeled sirna is particularly wellsuited for use in doublelabe. Quantitative comparison of longwavelength alexa fluor dyes to cy dyes. Sulfocy5 nhs ester is a sulfonated, hydrophilic and highly water soluble dye. Significant sequence dependence is seen for both cy3 and cy5modified oligonucleotides. Live cell imaging lipophilic carbocyanine membrane dye suitable for longterm fluorescent cell labeling and cell. Also see our cellular stains comparison guides to browse targets, applications, fixability, and stains for different organisms at a glance.

These handy labels will help you create an efficient labeling system in record time. Amersham cydye dige fluor labeling kit for scarce samples price. Cyanine dyes cy2, cy3, and cy5 jackson immunoresearch. But the most popular cyanine dyes for life science research were introduced by alan waggoner and colleagues of carnegie mellon university in early 1990s. Selective labeling of cellsurface proteins using cydye dige fluor. Cydye dige fluor preparative gel labeling for scarce samples, cy3 saturation dye 400nmol cydye dige fluor minimal dyes and saturation dyes are size and chargematched fluorescent dyes specifically designed for detecting protein abundance differences in 2d fluorescence difference gel electrophoresis using ettan dige system. Cydye postlabelling reactive dye pack ge healthcare. Cydye dige fluor preparative gel labeling for scarce samples. Cy5 protein labeling kit, fluorescent amine protein labeling.

The results showed specific labeling of a large number of cell surface proteins with minimal labeling of intracellular proteins. Specifically optimized for nucleic acid microarray labeling. A method to map changes in bacterial surface composition. Labeling densities are optimized for each particular application. Selective labeling of cellsurface proteins using cydye dige fluor minimal dyes. Invitrogen cy3 dye is a bright, orangefluorescent dye that can be excited using the 532 nm laser line and visualized with tritc tetramethylrhodamine filter sets. Selective labelling of cellsurface proteins using cydye dige fluor minimal dyes. The key feature of the dige method relies on labeling of protein samples. Cyanine is the nonsystematic name of a synthetic dye family belonging to polymethine group. Gel electrophoresis dige utilizing cyanine dyes cy dyes. Unexpectedly, the optimal dyeprotein ratio strongly depends on the type of cy dye. Among currently available fluorescent dyes, the cyanine dyes are better able to withstand the harsh dehydration and embedding conditions required for mounting sections in nonpolar plastic media, such as dpx and permount.

Selective labeling of cellsurface proteins using cydye dige. This reagent is specially quantified each package contains specified amount of nhs ester with quantity variation within 10%. Soluble in aqueous labeling media as the activated nhs ester, reducing precipitation that leads to inefficient protein labeling. Invitrogen cy5 dye is a bright, farredfluorescent dye with excitation ideally suited for the 633 nm or 647 nm laser lines. Selective labeling of cellsurface proteins using cydye. Hook 550 dyenhs ester is a fluorescent dyenhs ester used for labeling antibodies, proteins, nucleic acids for application is biochemical detection assays including flow cytometry western blotting, microscopy and imaging. For larger quantities of dye please order 25800861 10nmol and 25190028 25nmol the dye is dissolved in dimethylformamide dmf and the minimal labeling reaction is quenched with llysine. Cydye dige fluor minimal labeling kit ge healthcare, 25801065, pack of 5 nmol. Lick your labeling problems with smead labeling software solutions or traditional selfadhesive labeling products. Scanning cydye dige fluor saturation dye gels using typhoon variable mode imager 51. Hot asymmetric pcr katze lab in vitro transcription.

Atto dyes and tracy dyes for fluorescent protein labeling. Cy5 dye is compatible with various instrumentation including many fluorescent microscopes, imagers, scanners, and fluorescence readers. If cy dye is attached to a pyridine nucleotide, it tends to stabilize the duplex more. This regent is especially useful for the labeling of proteins which denaturate in the presence of organic cosolvents, as well as for proteins with low solubility. Label it nucleic acid labeling kits frequently asked questions. Cydye dige fluor minimal dyes and saturation dyes are size and chargematched fluorescent dyes specifically designed for detecting protein abundance. Printing your own labels is a good way to cut costs, but youll need the. This jena bioscience protein labeling kit is designed for labeling the lysines of a poi with a small fluorophore resulting in a fluorescent proteinfluorophore conjugate. Chemical safetys online cloudbased ghs labeling software allows you to find, display, and print ghs compliant labels in a multitude of sizes and languages. I know now that ge makes the monofunctional dye packs, but what i am interested in is to buy the chemicals as a powder form. For a more comprehensive list of citations to this article, users are encouraged to perform a search inscifinder. Labeling was done with cyanine 2, 3 and 5 cy 2, cy 3 and cy 5, respectively.

The distinct signal from each cydye label plays a part in the high. Osha mandates the labeling of chemical products with ghs safety information to highlight hazards and dangers. Lgc biosearch technologies series of quasar dyes are direct, costeffective replacements for the cy 3, cy5 and cy5. I would like to know the companies where you buy the bulk amount from.

For the labeling of various aminecontaining molecules in aqueous phase, no organic cosolvent is needed. See q for details about calculating the labeling efficiency of your sample. Using the general label it nucleic acid labeling kits, we routinely achieve one label every 2060 base pairs estimated with a spectrophotometric assay for the fluorescent label it dyes and dnp. Selective labelling of cellsurface proteins using cydye dige fluor. Features and benefits the only reactive cydye fluors specifically optimized for nucleic acid microarray labeling. Hook 550 dyenhs ester is a water soluble amine reactive. Cy3, tetramethylrhodamine tritc is a bright, orangefluorescent dye that can be excited by mercury arc lamps using the 532 nm laser line and detected with tritc tetramethylrhodamine filter sets. Ambion labeled sirna can be used to analyze sirna subcellular localization, stability, and transfection efficiency.