Flamma® 675 NHS ester

Product#: PWS1515

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Flamma® 675 NHS ester

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Flamma® Fluors 675 NHS ester is a reactive form of near infrared (NIR) fluorescent dye induced from benzindocyanine structure and used to generate a stable fluorescence signal in bioimaging. The maxima of Ex/Em values are at 675/691 nm, similar to that of Alexa 680, Cy5.5, IRDye 680LT and Dylight 680. Flamma 675 might be excited using the 633 nm laser line and the emission occurs at biological tissue permeable NIR region. Flamma 675 dye can be conjugated to low-abundance biomolecules in fixed cells with great sensitivity, and NIR emission allows in vitro imaging in biological studies. NHS esters readily react with amine-modified oligonucleotides or amino groups of proteins, i.e. the ε-amino groups of lysine or the amine terminus of nucleotides to form a chemically stable amide bond between dye and the biomolecule. We offer Flamma Fluors 675 dye for labeling of antibodies, peptides, proteins, ligands, and amplification substrates optimized for in vitro imaging.
  • Fluorophore: Flamma® Fluors 675
  • Reactive group: NHS ester
  • Excitation/Emission Max.(nm): 675/691 
  • Spectrally similar dyes: Alexa680, DyLight680, Cy5.5, IRDye680LT, CF680
  • Extinction coefficient: ≥ 200,000 cm-1M-1
  • CF280: 0.09
  • Appearance: Blue Solid
  • Molecular Weight: 1042.18 g/mol   
  • Solubility: DMF, DMSO
  • Storage conditions: -20 ℃, protect from light
Quick link (Cat.#) FSD series EXmax (nm) EMmax (nm) Spectrally similar dyes
CWS1001 Flamma® 496 NHS ester 496 520 Alexa488, FITC, Cy2
PWS1122 Flamma® 552 NHS ester 550 564 Alexa555, DyLight549, Cy3, ATTO550
PWS1415 Flamma® 581 NHS ester 581 596 Alexa594, DyLight594
PWS1215 Flamma® 648 NHS ester 648 663 Alexa647, DyLight650, Cy5
PWS1515 Flamma® 675 NHS ester 675 691 Alexa680, DyLight680, Cy5.5, IRDye680LT, CF680
PWS1301 Flamma® 749 NHS ester 749 774 Alexa750, DyLight755, Cy7.5, IRDye750
PWS1603 Flamma® 774 NHS ester 774 800 Cy7.5, DyLight800, IRDye800


Flamma® Fluors

BioActs offers a broad range of Flamma® Fluors dyes equipped with variety of reactive and functional groups, which can cover the full spectral range from UV to NIR with their excellent fluorescence performance. Characteristic features of these superior dyes are strong absorption, high fluorescence quantum yield and high photostability. Flamma® dyes maintain good fluorescence activity and stability after conjugation to biomolecules and allow the detection of low-abundance biological structures with great sensitivity. The dyes are compatible with optical conditions of most of fluorescent equipment and are ideal for any applications in biological studies.
  • Covering the full spectral range from UV to NIR
  • Equipped with a variety of reactive groups: NHS and Sulfo-NHS ester, Vinylsulfone, Maleimide, Click chemistry, isothiocyanate, hydrazide and hydrophobic substances.
  • High quantum yields and photostability
  • High purity and compatible with most of biomolecules


Flamma® Fluors NHS ester / Sulfo-NHS ester

Succinimidyl (NHS) esters are reliable activating moiety for amine labeling due to the formation of the strong amide bond. NHS esters are generally stable when storing in anhydrous and the dark condition at -20. NHS ester dyes display good reactivity toward aliphatic amines yet low reactivity with aromatic amines, alcohols, phenols and histidine. The basicity of an amine is an important factor for its reactivity: virtually all proteins have lysine residues, and most of them have a free N-terminus amine. The reactivity of ε-amino group at lysine is pH dependent that the optimal pH for labeling lysine residue is 8.5~9.5. However, the primary amine at N-terminus usually has a pKa value of ~7, thus it can be selectively conjugated by running the reaction in neutral pH. Due to the solubility issue, Flamma® Fluors NHS esters are required to be dissolved in an organic solvent such as DMSO or DMF and added to aqueous solution. To overcome this limitation, BioActs offers water soluble Flamma® Fluors Sulfo-NHS ester dyes, which can eliminate the usage of organic solvents in the conjugation reaction. NHS and Sulfo-NHS ester dyes are unstable under humid condition, thus they should be in a lyophilized form or a solution in the anhydrous organic solvent such as DMF or DMSO and stored at -20 ℃.


Figure 1.   Absorption (upper) and emission (bottom) spectra overlap of Flamma® Fluors


Figure 2.   Immunofluorescence imaging and in situ hybridization imaging


Figure 3.   Fluorescence images of Flamma® 749 (upper) and Flamma® 774 (bottom) carboxylic acid injected mouse

Citation & Reference

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