Which BV dyes are tandem?

BioLegend available tandems

Tandem	Exmax(nm)	Bandpass Filter
Brilliant Violet 605™	405	610/20
Brilliant Violet 650™	405	660/20
Brilliant Violet 711™	405	710/50
Brilliant Violet 750™	405	780/60

What are tandem dyes in flow cytometry?

Tandem dyes refer to compounds in which two different fluorophores have been covalently linked. One of the fluorophores serves as donor and the other as acceptor, forming a fluorescence-resonance energy transfer (FRET) pair.

What is a tandem fluorochrome?

A tandem dye, also called a FRET pair or FRET dye, is a pair of covalently linked fluorescent molecules (fluorochromes or fluorophores) which contain a donor and an acceptor molecule (Figure 1A).

Are Alexa Fluor tandem dyes?

Additionally, we have conjugated allophycocyanin to Alexa Fluor 680, Alexa Fluor 700 or Alexa Fluor 750 dyes to create tandem conjugates for multicolor measurements using excitation sources in the 633 to 650 nm range (Figure 1.3. 25).

Are brilliant violet tandem dyes?

The six Brilliant Violet polymer dyes come in two forms—a base polymer and a tandem polymer. These dyes are very bright and in many cases, provide unprecedented brightness for dyes excited by the violet laser (Figure 1).

Is BV480 a tandem?

About Brilliant Violet 480 Brilliant Violet™ 480 (BV480) is a green-emitting non-tandem polymer fluorophore that can be excited by the 405 nm violet laser and collected using a 525/40 bandpass filter.

Why are tandem dyes used?

Tandem dyes largely expand the pool of fluorophores that are suitable for flow cytometry, increasing the number of colors that can be detected in a single experiment. The number of fluorophores one can use at the same time is limited by the number of lasers and detectors on a flow cytometer.

What types of fluorochromes exist?

In general, fluorochromes can be divided into 5 broad categories, which are discussed below.

• Fluorescent Proteins. Fluorescent proteins can be categorized into two groups.
• Synthetic Small Molecules.
• Quantum Dots.
• Polymer Dyes.
• Tandem Dyes.

Can I use PE and PE Cy7 together?

In my experience, it is not advisable to use APC/Cy7 and PE/Cy7 together due to heavy cross-beam contamination as their emission max are exactly same. However, they can be used together if you are using machines like the Cytek Aurora.

Is Alexa Fluor 647 a tandem dye?

This enables multicolor analysis in conjunction with APC or Alexa Fluor® 647 and APC-H7 or APC-Cy7 reagents. 633 nm 635 nm 640 nm 780/60 APC-Cy™7 (Exmax650 nm/Emmax785 nm) is a tandem fluorochrome that combines APC and the cyanine dye Cy7.

What type of fluorochromes exist?

In general, there are two classes of fluorochrome used in flow cytometry – those which bind non-covalently to structures within the cell and those which are covalently bound to other probes. The fluorescent proteins, such as Green Fluorescent Protein, (GFP) form a special category.

What is the primary advantage of tandem dyes over conventional fluorescent dyes?

Can I use PE and APC together?

488 nm 532 nm 561 nm 670/14 PE-Cy™5 (Exmax496 nm/Emmax667 nm) is a tandem conjugate that combines phycoerythrin and the cyanine dye Cy5. Because of its broad absorption range and the fact that its emission spectra are equivalent to APC, PE-Cy5 is not recommended for simultaneous use with APC.

Is BB700 a tandem?

About BB700 Horizon Brilliant™ Blue 700-P (BB700-P) is a tandem dye member of the BD BioSciences Horizon Brilliant Blue family of dyes.

Is BV421 fixable?

However, BV421™ is consistently demonstrated to be compatible with viability probes Fixable Live/Dead Blue or DAPI excited off the UV laser.

Is PE Cy7 bright?

PE-Cy7 conjugated reagents are as bright as PE conjugates. However, PE-Cy7 is highly sensitive to photo-induced degradation resulting in loss of fluorescence. Care should be taken to avoid light exposure and prolonged exposure to paraformaldehyde fixative.

Can I use APC and PE together?

PerCP overlap a bit with APC and PeCy7. Pe-‐CF594 spill over a lot from and to Pe, don’t use together. PeCy5 spill over a lot to APC (bad), Pe and PeCy7. Avoid these combinations if possible.

Is 647 the same as Cy5?

For stable signal generation, Alexa Fluor 647 dye is generally brighter than Cy5 dye in protein and nucleic acid conjugates. Allophycocyanin (APC) is another bright fluorophore that is spectrally equivalent to Cy5 dye and Alexa Fluor 647 dye, but it is used almost exclusively in flow cytometry.

What is the difference between FACS and flow cytometry?

FACS is used as a cell sorter and enriched for a subset of cells which is often then studied in further detail using flow cytometry or other analytical techniques2. Flow cytometry is used for cell analysis and is focused on measuring protein expression or co-expression within a mixed population of cells.

Is APC brighter than PE?

It is also because APC is red-shifted compared to PE. Thus, the light source used to maximally excite APC will contain less energy than the one used to maximally excite PE. Compared to other red fluorophores such as Cy5, however, APC is still significantly brighter and an excellent choice for use in flow cytometry.

What is a tandem dye in fluorochromes?

Fluorochromes have unique and characteristic spectra for absorption (excitation) and emission. A single dye is excited at a particular wavelength and emits a photon at a higher wavelength. A tandem dye consists of a the two.

What are the characteristics of fluorochromes?

Fluorochromes have unique and characteristic spectra for absorption (excitation) and emission. A single dye is excited at a particular wavelength and emits a photon at a higher wavelength. A tandem dye consists of a

What is the brightest fluorochrome?

PE has one of the largest absorption co-efficiencies making it one of the brightest fluorochromes available to date. PE-Texas Red (Ex-Max 496 nm/Em-Max 615 nm): is a tandem conjugate that combines PE and Texas Red.

What is the peak absorbance of fluorochromes?

For example, the most widely used fluorochrome, Fluorescein isothiocyanate (FITC), has a peak absorbance of 494nm and a peak emission of 512nm corresponding to the colors blue and green. Exciting fluorochromes at their peak absorbance (rather than at the bottom end of their absorbance spectrum), enables more intense light emission.