"I have used a wide variety of secondaries and Jackson ImmunoResearch has consistently been the best. The fluorophores are bright and stable and their selective (x reactivity removed) secondaries have always shown species specificity in multiple labeling."
Janet Duerr, Ohio University
Brilliant Violet 421 and Brilliant Violet 480 conjugated secondary antibodies allow you to add more colors to your multiple labeling assays in the violet-blue region of the spectrum.
Double immunofluorescence of Hep-2 cells using Brilliant Violet 421 and 480 conjugated secondary antibodies. Nuclei counterstained with DRAQ5.
Add more colors to multiple labeling assays with Brilliant Violet 421™ and Brilliant Violet 480™ conjugated secondary antibodies. When combined with Alexa Fluor® 488, Rhodamine-Red™-X, and Alexa Fluor® 647 conjugates, effective 5-color fluorescent labeling is possible.
If nuclear counterstaining is desired, 4-color antibody staining is possible using Brilliant Violet 421, Brilliant Violet 480, Alexa Fluor® 488 and Rhodamine Red™-X. Switching the nuclear stain from DAPI (emission in the blue region) to DRAQ5™ (which has red emission) frees the violet-blue region of the spectrum to accommodate the two Brilliant Violet dyes. DRAQ5’s excitation and emission profiles overlap those of Alexa Fluor 647.
These three options are summarized in the table below.
| Channel | Blue | Cyan | Green | Orange | Red |
| Option 1 | DAPI | BV480 | Alexa Fluor 488 | RR-X | Alexa Fluor 647 |
| Option 2 | BV421 | BV480 | Alexa Fluor 488 | RR-X | DRAQ5 |
| Option 3 | BV421 | BV480 | Alexa Fluor 488 | RR-X | Alexa Fluor 647 |
We offer a range of secondary antibodies that are recommended for multiple labeling due to their minimal cross-reactivity, ( See our multiple labeling guidelines for more information).
Available Target Specificities for Multiple Labeling (Designated ML* in product tables)
Browse Brilliant Violet™ Secondary Antibody Conjugates
| Excitation Peak (nm) | Emission Peak (nm) | |
|---|---|---|
| Brilliant Violet 421™ | 407 | 421 |
| Brilliant Violet 480™ | 436 | 478 |
Excitation and emission curves for BV 421™, BV 480™, Alexa Fluor ® 488, Rhodamine Red™-X, and Alexa Fluor® 657.
DRAQ5 can be used as a nuclear stain in place of Alexa Fluor 647.
BD Brilliant Violet polymer chains can be considered as a collection of optical segments, each with the ability to absorb light and emit fluorescence signal. This results in materials that have a bright fluorescence signal for better resolution and sensitivity.
Light harvesting base polymers have multiple optical segments that can emit for fluorescence detection.
| Channel | Fluorophore | Excitation (nm) | Emission (nm) | Chroma Filter Set |
|---|---|---|---|---|
| Blue | BV421™ | 407 | 421 | 49027 |
| Cyan | BV480™ | 439 | 477 | 49302 or 49001 |
For more information on these filter sets, please go to the Chroma website.
| Fluorophore | Extinction co-efficient (M -1 cm-1) | Quantum Yield (ɸF) |
|---|---|---|
| BV421™ | 2587000 | 0.91 |
| BV480™ | 1416000 | 0.81 |
Brilliant Violet™ is a trademark of Sirigen Inc., a Becton, Dickinson and Company affiliate.
