Alexa Fluor® 555 and 568 secondary antibody conjugates are now available from Jackson ImmunoResearch. With the addition of two new choices for the orange channel, panel design has become even easier.
| Alexa Fluor®-Dye | Excitation (nm) | Emission (nm) |
|---|---|---|
| Alexa Fluor® 488 | 493 | 519 |
| NEW: Alexa Fluor® 555 | 552 | 572 |
| NEW: Alexa Fluor® 568 | 577 | 602 |
| Alexa Fluor® 594 | 591 | 614 |
| Alexa Fluor® 647 | 651 | 667 |
| Alexa Fluor® 680 | 684 | 702 |
| Alexa Fluor® 790 | 792 | 803 |
Table 1: Alexa Fluor® dyes available conjugated to Jackson ImmunoResearch products.
Figure 1: Excitation and emission spectra for Alexa Fluor® dyes available conjugated to Jackson ImmunoResearch products (Fig. curtesy of Jackson ImmunoResearch).
Alexa Fluor® 555 and Alexa Fluor® 568 dyes are available conjugated to donkey and goat secondary antibodies and allow multiplex labeling with minimal cross-reactivity to many common species.
Alexa Fluor® fluorophores are widely recognized as the superior fluorophores available for conjugation. They are highly soluble in water and remain fluorescent from a pH of 4 to a pH of 10.
The detection level of any fluorophore-antibody conjugate is dependent on the brightness and photostability of the dye; the activity, specificity, and cross-reactivity of the antibody; and the optimal number of moles of dye per mole of antibody. These parameters have been optimized by Jackson ImmunoResearch for each dye conjugate to offer the best antibody detection levels and minimize background.
Properties of Jackson ImmunoResearch Alexa Fluor® 555 Secondary Antibodies
| Alexa Fluor®-Dye | Laser-Line (nm) | Excitation (nm) | Emission (nm) | Extinktionskoeffizient (M -1 cm-1) | Quantum Yield(ɸF) | Molecular Weight (kDa) | Applications |
|---|---|---|---|---|---|---|---|
| Alexa Fluor® 555 | 488/532 | 493 | 519 | 1.4 | 0.1 | 1250 | Microscopy, Flow Cytometry, Fluorisa, Fluorescent Western Blotting |
Properties of Jackson ImmunoResearch Alexa Fluor® 555 Secondary Antibodies
Alexa Fluor® 555 conjugated antibodies absorb light maximally at 552 nm and fluoresce with a peak at 572 nm. Alexa Fluor® 555 Dye is a bright, orange fluorescent dye whose excitation is ideally matched to the 555 nm laser line. Alexa Fluor® 555 dye is pH-insensitive over a wide molar range, providing stable signal generation for flow cytometry and imaging applications, including some super-resolution protocols (Goossen-Schmidt et al., 2020). Dye molecules such as Alexa Fluor® 555 with high fluorescence quantum yield and high photostability enable the detection of low-abundance biological structures with high sensitivity.
Properties of Jackson ImmunoResearch Alexa Fluor® 568 Secondary Antibodies
| Alexa Fluor®-Dye | Laser-Line (nm) | Excitation (nm) | Emission (nm) | Extinktionskoeffizient (M -1 cm-1) | Quantum Yield(ɸF) | Molecular Weight (kDa) | Applications |
|---|---|---|---|---|---|---|---|
| Alexa Fluor® 568 | 568 | 577 | 602 | 1.4 | 0.1 | 792 | Microscopy, Flow Cytometry, Fluorisa, Fluorescent Western Blotting |
Properties of Jackson ImmunoResearch Alexa Fluor® 568 Secondary Antibodies
Alexa Fluor® 568 conjugated antibodies absorb light maximally at 577 nm and fluoresce with a peak at 602 nm. Alexa Fluor® 568 dyes are bright, orange fluorescent dyes with excitation that is ideal for the 568 nm laser line of the Ar-Kr mixed gas laser. Alexa Fluor® 568 dyes are pH insensitive over a wide molar range. Alexa Fluor® 568 is a bright and photostable orange-red dye that works well with Red Fluorescent Protein (RFP) filter sets and can be used in Stochastic Optical Reconstruction Microscopy (STORM) – as a reporter in dSTORM (Goossen-Schmidt et al., 2020).