Molecular Probes™ Alexa Fluor™ Carboxylic Acid, tris(triethylammonium) salts

Description

• Alexa Fluor Carboxylic Acid tris(triethylammonium) salts are non-reactive, free acid forms of Alexa Fluor dyes
• These carboxylic acid derivatives can be converted to an amine-reactive ester using standard chemical techniques or coupled to hydrazines, hydroxylamines, and amines in aqueous solution using EDAC
• Discover the benefits of the Alexa Fluor dye series Compared to traditional dyes, Alexa Fluor dyes are brighter, more photostable, and more pH-resistant between pH 4 and 10
• Generally when using Alexa Fluor dyes, higher degrees of labeling can be achieved without intramolecular quenching
• Thermo Fisher Scientific offers a variety of free acid dyes using carboxylic acid tris(triethylammonium) salts to provide non-reactive forms of various Alexa Fluor dyes
• These Alexa Fluor dyes are offered as a free acid, where you can choose from ten different colors and wavelength excitations across the fluorophore spectrum
• Being unconjugated and non-reactive, Alexa Fluor free acid dyes cannot be used for labeling
• However, Alexa Fluor free acid molecules are useful for injection site visualization in vivo and can be used as reference standards for corresponding dye-conjugates in any fluorescent labeling experiment
• Free acid formats of Alexa Fluor dyes can be useful controls for confocal imaging, ICC/IHC, HCS, flow cytometry and live cell imaging
• To reconstitute, dissolve the Alexa Fluor carboxylic acid compound in aqueous buffer (e.g
• PBS), or high-quality, anhydrous dimethylsulfoxide (DMSO) or dimethylformamide (DMF) at a final concentration of 1–10 mg/mL
• Once reconstituted, this solution is stable for 1 year, when stored at 2–8°C and protected from light
• Alexa Fluor free acid dye options: • Blue-fluorescent Alexa Fluor 350 —excitation and emission maxima of 346/442 nm; excited using a 355-nm laser line and detected under standard DAPI filters
• Alexa Fluor 350 is commonly used with Alexa Fluor 488, 594, and 647 dyes for multiplexing
• • Green-fluorescent Alexa Fluor 488 —excitation and emission maxima of 490/514 nm; excited using a 488-nm argon laser line and detected under standard FITC/Cy2 filters
• Alexa Fluor 488 is commonly used with DAPI, Alexa Fluor 594 and 647 dyes for multiplexing
• • Yellow-green fluorescent Alexa Fluor 532 —excitation and emission maxima of 528/548 nm; excited using a 532-nm Nd:YAG laser line and detected under standard rhodamine 6G filters
• Alexa Fluor 532 free acid dye has excitation and emission spectra between those of the green-fluorescent Alexa Fluor 488 dye and the orange-fluorescent Alexa Fluor 546 dye
• • Yellow-fluorescent Alexa Fluor 546 —excitation and emission maxima of 554/570 nm; excited using a 543-nm He-Ne laser line and detected under standard TRITC/Cy3 filters
• Alexa Fluor 546 serves as an alternative for tetramethylrhodamine or Cy3 dye
• • Orange-fluorescent Alexa Fluor 555 —excitation and emission maxima of 555/572 nm; excited using a 543-nm He-Ne laser line and detected under standard TRITC/Cy3 filters
• Alexa Fluor 555 is a recommended replacement for Cy3 dye or tetramethylrhodamine, with greater photostability than Cy3
• Alexa Fluor 555 dye is commonly used with DAPI, Alexa Fluor 488 and 647 dyes for multiplexing
• • Orange-red-fluorescent Alexa Fluor 568 —excitation and emission maxima of 571/596 nm; excited using a 568-nm Ar-Kr mixed-gas laser and detected under standard RFP/Rhodamine Red/Cy3.5/Texas Red filters
• Alexa Fluor 568 dye is commonly used with DAPI, Alexa Fluor 488 and 647 dyes for multiplexing
• • Red-fluorescent Alexa Fluor 594 —excitation and emission maxima of 585/610 nm; excited using a 594-nm Kr or He-Ne laser line and detected under standard Texas Red filters
• Alexa Fluor 594 is a direct replacement for Texas Red dye with bright signal and excellent photostability
• Alexa Fluor 594 dye is commonly used with DAPI, Alexa Fluor 488 and 647 dyes for multiplexing
• • Deep-red-fluorescent Alexa Fluor 633 —excitation and emission maxima of 620/640 nm; excited by the 633 nm spectra line of the He-Ne laser or the 635 nm spectral line of red diode lasers
• This and other far-red fluorescent Alexa Fluor dyes are important for fluorescence imaging because their spectra are beyond the range of most sample autofluorescence
• • Deep-red-fluorescent Alexa Fluor 647 —excitation and emission maxima of 651/672 nm; excited using a 633- or 635-nm Kr or He-Ne laser line and detected under standard APC/Cy5 filters
• Alexa Fluor 647 dye is commonly used with DAPI, and Alexa Fluor 488 and 594 dyes for multiplexing
• • Near-IR Alexa Fluor 750 —excitation and emission maxima of 753/783 nm; excited using 750 nm laser line and detected under standard Cy7 filters
• Alexa Fluor 750 is well separated from commonly used far-red fluorophores such as Alexa Fluor 647 or APC
• Carboxylic acid derivatives can be converted to an amine-reactive ester using standard chemical techniques or coupled to hydrazines, hydroxylamines, and amines in aqueous solution using EDAC (Cat
• No
• E2247)
• EDAC is a water-soluble carbodiimide that can be used to crosslink biological substances that contain carboxylate acids and primary amines.