18
2.
Monolith
®
NT.115 Instrument
2.1. Models and Most Common Fluorophores
Fluorophore
Excitation
[nm]
NT.115
Blue/Green
NT.115
Blue/Red
NT.115
Green/Red
NT.115
Pico
Emission
[nm]
BCECF
480
525
GFP
488
507
NT-495 (BLUE)
493
521
Fluorescein (FITC)
495
519
Alexa488
495
519
YFP
514
527
Alexa532
530
555
TAMRA
546
579
Cy3
550
570
RFP
555
584
NT-547 (GREEN)
557
574
Alexa546
560
572
Cy5
649
670
NT-647 (RED)
650
670
Alexa647
652
668
Note: Excitation and emission wavelengths can vary depending on the environment (buffer, pH).
Colors are only a very rough approximation. The minimum concentration for use in NT.115 series
instruments can vary.
2.2. Application Range
The Monolith NT.115 allows measurement of the binding of virtually all biomolecules over a large
range of binding affinities (
K
d
= 1 nM - 500 mM).
2.3. Sensitivity
The Monolith NT.115 can measure as little as the binding of single ions (40 Da) or small molecules
(300 Da) to a target as well as the binding of large complexes such as ribosomes (2.5 MDa).
2.4. Sample Consumption
The Monolith NT.115 requires minimal sample material: typically 1 -10 nM of the labeled interaction
partner and a dilution series of the unlabeled ligand starting about 20-fold above the expected
dissociation constant. For standard applications, 4
μl of sample material is loaded in the capillary.
