Create mode – the default mode when you create a requisition and PunchOut to Bio-Rad. You can create and edit multiple shopping carts
Edit mode – allows you to edit or modify an existing requisition (prior to submitting). You will be able to modify only the cart that you have PunchedOut to, and won't have access to any other carts
Inspect mode – when you PunchOut to Bio-Rad from a previously created requisition but without initiating an Edit session, you will be in this mode. You cannot modify any Cart contents
Bio-Rad-Antibodies.com relies on third-party cookies to show you pricing, allow you to order online, and connect you to My Bio-Rad. Please amend your browser settings to enable third-party cookies and access this website’s full functionality.
There are many fluorescent molecules (fluorophores) with a potential application in flow cytometry. The list is ever growing and we will not cover all of them here. The fluorophores currently available from bio-rad-antibodies.com are described in Tables 1 and 3. With new fluorophores constantly being added, don’t forget to check out our latest flow cytometry products.
Single dyes such as FITC, PE, APC and PerCP have been available for many years, but there are now alternatives available from Alexa Fluor dyes, which offer users greater photostability and brighter fluorescence. In addition alternative laser lines are becoming more affordable so dyes excited by 355 nm and 405 nm lasers are increasing the options for multiplexing.
Tandem dyes comprise a small fluorophore covalently coupled to another fluorophore. When the first dye is excited and reaches its maximal excited electronic singlet state, its energy is transferred to the second dye (an acceptor molecule). This activates the second fluorophore, which then produces the fluorescence emission. The process is called Förster resonance energy transfer (FRET). It is a clever way to achieve a higher Stokes shift and therefore increase the number of colors that can be analyzed from a single laser wavelength.
The majority of tandem dyes have been manufactured for the 488 nm and 640 nm lasers which are found in most cytometers. Tandem dyes are very useful for multicolor fluorescence studies, especially in combination with single dyes. For example Alexa Fluor 488, phycoerythrin (PE), peridinin chlorophyll protein (PerCP)-Cy5.5 and PE-Texas Red can all be excited at 488 nm, but will produce green, yellow, red and infrared emissions, respectively, which can then be measured using separate detectors.
Fluorescent proteins, such as green fluorescent protein (GFP), have become an integral tool for understanding protein expression in many scientific disciplines. Other fluorescent proteins, such as mCherry and yellow fluorescent protein, have also become widely used for flow cytometry analysis and cell sorting. The fluorescent proteins are often co-expressed or expressed as a fusion with the protein of interest. The benefit of these fluorescent proteins is the quantitation of intracellular markers in live cells without requiring permeabilization of the cell membrane. Common fluorescent proteins are listed in Table 4.
Fluorophores | Fluorescence Color | Maximum Excitation (nm) | Maximal Emission (nm) | Relative Brightness |
---|---|---|---|---|
DyLight 405 | 400 | 420 | 3 | |
Alexa Fluor 405 | 401 | 421 | 3 | |
Pacific Blue | 410 | 455 | 1 | |
Alexa Fluor 488 | 495 | 519 | 3 | |
FITC | 490 | 525 | 3 | |
DyLight 550 | 562 | 576 | 4 | |
PE* | 490; 565 | 578 | 5 | |
APC | 650 | 661 | 4 | |
Alexa Fluor 647 | 650 | 665 | 4 | |
DyLight 650 | 654 | 673 | 4 | |
PerCP | 490 | 675 | 2 | |
Alexa Fluor 700 | Infrared | 702 | 723 | 2 |
* PE is the same as R-phycoerythrin.
APC, allophycocyanin; FITC, fluorescein isothiocyanate; PE, phycoerythrin; PerCP, peridinin chlorophyll protein.
Fluorophores | Fluorescence Color | Maximum Excitation (nm) | Maximum Emission (nm) | Relative Brightness |
---|---|---|---|---|
StarBright Violet 440 | 383 | 436 | 5 | |
StarBright Violet 515 | 402 | 516 | 5 | |
StarBright Violet 610 | 402 | 607 | 5 | |
StarBright Violet 670 | 401 | 667 | 5 | |
StarBright Blue 700 | 470 | 705 | 5 |
Fluorophores | Fluorescence Color | Maximum Excitation (nm) | Maximal Emission (nm) | Relative Brightness |
---|---|---|---|---|
PE-Alexa Fluor® 647 | 496, 546 | 667 | 4 | |
PE-Cy5 | 496, 546 | 667 | 5 | |
PerCP-Cy5.5 | 490 | 695 | 3 | |
PE-Cy5.5 | 496, 546 | 695 | 4 | |
PE-Alexa Fluor® 750 | Infrared | 496, 546 | 779 | 4 |
PE-Cy7 | Infrared | 496, 546 | 785 | 2 |
APC-Cy7 | 650 | 785 | 2 |
* PE is the same as R-phycoerythrin.
APC, allophycocyanin; PE, phycoerythrin; PerCP, peridinin chlorophyll protein.
Fluorophores | Fluorescence Color | Maximum Excitation (nm) | Maximal Emission (nm) | Relative Brightness |
---|---|---|---|---|
EGFP | 383 | 445 | 2 | |
CFP | 439 | 476 | 2 | |
EGFP | 484 | 509 | 4 | |
YFP | 514 | 527 | 5 | |
RFP | 558 | 583 | 4 | |
mCHERRY | 587 | 610 | 3 |