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Detection

Atto

Atto labels represent a comprehensive series of fluorescent labels for biomolecules, which are designed for highest sensitivity applications. A unique combination of advantages makes them highly favorable tools for all kinds of labeling applications.
Features and Benefits
Spectral data: Absorption maxima fit commonly used excitation sources. Their emission maxima cover a broad range from 500 to 700 nm.
Stokes' shift: Shift between absorption maxima and emission maximum is sufficient to discriminate the fluorescence emission from scattered excitation light.
Brightness: Bright fluorescence due to strong absorbance (molar extinction above 10 5) and high quantum yields (ranging from 0.3 to 0.9).
Photostability: Structural properties support an extraordinary photostability.
Hydrophilicity: Good, supporting labelling in aqueous solutions.
Reactivity: All dyes are available as activated NHS-esters, matching most common labelling procedures for proteins and amino functionalized oligonucleotides. Besides that, a series of thiol reactive maleimides is available.
Atto_Stability
The absence of cis-trans-isomerization circumvents potential problems in Fluorescent Resonant Energy Transfer (FRET) or single molecule detection (for more details and references please see Analytix 8/2001)
Our range of labels show excitation and emission maxima spanning the range from 390 - 750 nm. This gives opportunity to use various sources of excitation light and to match emission light common used filter sets for other labels. Obviously it also allows a variation of multicolor visualization. The comparably small size of our labels (MW between 400 and 600) minimizes risk of unwanted steric effects towards the interaction of labeled molecules with other target structures. Furthermore Atto labels are quite insensitive against pH fluctuation - most of them do not show any effects between pH 2 and 10- and feature low unspecific adsorption.
spectrum1
spectrum2
Fig. 1: Absorption and emission spectra of Atto labels
Table 1: Properties of Atto dyes measured in water (ethanol) 

Product Name
e [M-1cm-1]
lmaxabs [nm]
lmaxem [nm]
h fl [%]
Atto 390 24'000  390  470 (475)   90 
Atto 425 45'000  436  484  90 
Atto 465 75'000  454  505  55 
Atto 488 105'000  504  521  80 
Atto 520 110'000 520 (525) 542 (547) 90 (95)
Atto 532 115'000 533 (560) 560 (560) 90 (90)
Atto 550 115'000 554 (556) 576 (578) 80 (90)
Atto 565 120'000 561 (566) 585 (590) 92 (97)
Atto 590 120'000 598 (598) 634 (634) 80 (90)
Atto 594 12'000 601 627 85
Atto 610 110'000 605 (616) 646 (630) 70 (70)
Atto 620 120'000 620 (620) 641 (646) 50 (70)
Atto 635 120'000 635 (637) 659 (660) 25 (45)
Atto 647 120'000 645 (648) 673 (669) 20 (40)
Atto 647N 150'000 644 667 65
Atto 655 110'000 665 (655) 690 (680) 30 (50)
Atto 680 110'000 680 (675) 702 (699) 30 (40)
Atto 700 120'000 504 714 25
Atto 725   725 752 10

 

Atto dyes are a series of fluorescent dyes, that provide all the crucial properties required for modern fluorescent technologies, such as fluorescence microscopy, flow cytometry, fluorescence in situ hybridisation (FISH), receptor binding assays, or enzyme assays.
endothelial_cells_atto532 
Confocal micrograph of human endothelial cells (HUVAC) visualizing vimentin by indirect staining; anti-Vimentin (rabbit) followed by Atto-532 (green) conjugated anti-rabbit-antibody (goat). Counterstaining of cell nucleus by DAPI (blue).
endothelial_cells_atto550_655
Triple staining of human endothelial cells (HUVAC): vWillebrand factor was visualized with Atto 550 labelled antibody (green), Cadherin with Atto 655 labelled antibody (red) and nucleus by DAPI (blue).
All Atto labels are available as free fluorophores (which might be coupled via free COOH-groups) or succinimidyl (NHS-) esters, which can be coupled to proteins easily. Coupling procedures may be found on our product information sheets.
For convenient use of fluorescent labels in common assay formats we now also offer Atto labels conjugated to specific proteins, inlcluding Atto antibody conjugates. Biotin- and Streptavidin conjugates are the most commonly used among them. The high affinity of streptavidin (like avidin) to biotin has been the basis for wide spread use of streptavidin conjugates as secondary detection reagents in microscopy, flow cytometry, immunoassays, blot analysis and nucleic acid hybridization methods. Streptavidin was chosen as it shows much lower unspecific binding than avidin. Our streptavidin conjugates basically show same spectral spattern than unbound labels. An interesting feature is that several of our streptavidin conjugates show low fluorescence, but strong enhancement of fluorescence after binding to biotin. This phenomenon helps to improve sensitivity in case, that some conjugate might be left adsorbed unspecifically.
Atto labels are also available as Maleimides, which open up an alternative method to amino labelling. Maleimides are suitable for coupling to thiol containing groups such as those contained in cysteine residues. The same holds for thiol groups introduced as modifiers during automated synthesis (of for example oligonucleotides).
fl_bio_new_intprof
Fluorescence intensity profile of single dye molecules (Atto 650) on a glass surface.
Product List(Standard Pack Size is 1 mg) 

Catalog No.
Name
Corresponding NHS-ester
Corresponding Maleimide
Streptavidin Conjugate
Biotin Conjugate
89313 Atto 390 89204 89740    
- Atto 425 16805 49349 09260 28616
50712 Atto 465 53404 55607    
41051 Atto 488 41698 28562   30574
70706 Atto 520 77810 16590 41469 01632
06699 Atto 532 88793 68499    
42424 Atto 550 92835 30730   28923
75784 Atto 565 72464 18507 56304 92637
70425 Atto 590 79636 39887 40709 43208
78493 Atto 610 93259 41061 56767 43292
92716 Atto 620 67351      
08968 Atto 635 43429      
97875 Atto 647 07376 41784   30700
04507 Atto 647N 18373 05316    
93711 Atto 655 76245 80661 02744 06966
94875 Atto 680 75999 04971 16630 55819
30674 Atto 700 16986 50611 - -
47156 Atto 725 93725 - - -
 

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