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"I have used a wide variety of secondaries and Jackson ImmunoResearch has consistently been the best. The fluorophores are bright and stable and their selective (x reactivity removed) secondaries have always shown species specificity in multiple labeling."
Janet Duerr, Ohio University
Technical Service e-mail
JIR offers a wide range of fluorescent conjugates, covering the most commonly used excitation sources and filter sets from blue to infrared emissions. The following information describes some of the characteristics of fluorescent dye families and where they are best employed. Information about individual dyes can be found by selecting the dye from the table below.
Further information about selecting a fluorophore for your application is available on the fluorophore selection page.
Alexa Fluor® fluorescent dyes are widely recognized as superior fluorescent dyes, respected for their brightness and photostability. They are highly water soluble and remain fluorescent from pH 4 to pH 10.
JIR offers two BD Brilliant Violet dyes, BV421 and BV480 (these dyes are named for their emission maxima, while many fluorophores are named for the excitation maxima). BV dyes are polymer chains and can be considered as a collection of optical segments, each with the ability to absorb light and emit fluorescence signal. This results in dyes that have a bright fluorescence signal for superior resolution and sensitivity.
Among currently available fluorescent dyes, the cyanine dyes are better able to withstand the harsh dehydration and embedding conditions required for mounting sections in non-polar plastic mounting media such as DPX and Permount™. The cyanine dyes are brighter in the non-polar environment than in aqueous media, resulting in reduced acquisition time compared with DyLight™ and Alexa Fluor® dyes, even though those dyes are brighter in aqueous mounting media.
Jackson ImmunoResearch offers 3 fluorescent proteins, Phycoerythrin (R-PE), Allophycocyanin (APC), and Peridinin-Chlorophyll-Protein (PerCP). R-PE and APC are light-harvesting phycobiliproteins found in red, blue-green and cryptomonad algae. Jackson ImmunoResearch offers R-PE in the form found in red macrophytic algae (seaweed). APC is isolated from the blue-green alga Spirulina. PerCP is a fluorescent peridinin-chlorophyll-protein complex isolated from dinoflagellates. R-PE, PerCP and APC can be excited by light over a wide range of the visible spectrum, are highly water soluble, have relatively low isoelectric points, and lack potentially sticky carbohydrates. It should be noted that the relatively high molecular weights of these fluorescent proteins may preclude their use in procedures requiring good penetration into cells and tissues. They are predominantly intended for surface labeling of cells for flow cytometry.
The following seventeen fluorescent probes are currently available from Jackson ImmunoResearch. Click on each fluorophore in the table for more information.
|Fluorophore||Excitation Peak||Emission Peak (nm)|
|Brilliant Violet 421™||407||421|
|Aminomethylcoumarin Acetate, AMCA||350||450|
|Brilliant Violet 480™||436||478|
|Alexa Fluor® 488||493||519|
|R-Phycoerythrin, R-PE||many, 488||580|
|Rhodamine Red™-X, RRX||570||590|
|Alexa Fluor® 594||591||614|
|Allophycocyanin, APC||many, 650||660|
|Alexa Fluor® 647||651||667|
|Peridinin-Chlorophyll-Protein, PerCP||many, 488||675|
|Alexa Fluor® 680||684||702|
|Alexa Fluor® 790||792||803|
Fluorescent probes or fluorophores (fluorescent dyes or proteins) are coupled to a secondary antibody or streptavidin to allow visualization of an analyte. Each fluorophore has its own spectral characteristics, with excitation and emission spectra particular to the molecule. Use the spectra viewer to build dye panels and compare the suitability of dyes for your application. Four dyes typically recommended for multiple labeling are pre-loaded, these can be changed as required.