fluorophorereference guide - bio-rad laboratories · iiiiiiii wavelength (nm) 350 400 450 500 550...
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400 nm
425 nm
450 nm
475 nm
500 nm
525 nm
550 nm
575 nm
600 nm
625 nm
650 nm
Laser Lines
ArI 488 nm
ArI 514 nm
DPSSL (µGreen) 532 nm
GHeNe 543 nm
YHeNe 594 nm
Red diode 635 nm
Broad UV Excitation290-365 nm
Emission FiltersLP = Long pass filterDF = Band pass filterSP = Short pass filter
555 nm555LP
VersaDoc FiltersMolecular Imager FX Filters
520 nm520LP
660 nm
515 nm
530DF30
545 nm
622.5 nm
640DF35
657.5 nm
580 nm
605DF50
630 nm
670 nm
690DF50
500 nm
530DF60
560 nm
610 nm610LP
660 nm
Excitation Maxima
Excel. ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ DAPI: 359 nm ____Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ GFP (Green Fluorescent Protein): 395 nm ____
Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Coumarin: 402 nm ____
Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ AttoPhos: 440 nm ____
Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Acridine Orange: 460/500 nm ____
Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ R-phycoerythrin: 480/565 nm ____Excel. ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ SYPRO Orange/SYPRO Ruby: 480 nm ____Excel. ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ NanoOrange: 485 nm ____Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Cy2: 489 nm ____Excel. ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ GelStar: 490 nm ____Excel. ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Oregon Green: 490 nm ____Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ FAM (5-FAM): 492 nm ____Excel. ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ SYBR Green II: 492 nm ____Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ FITC: 494 nm ____Excel. ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ SYBR Green I: 494 nm ____Med. ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Alexa Fluor 488: 495 nm ____Excel. ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Radiant® Red: 495 nm ____Excel. ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ SYBR Gold: 495 nm ____Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ SYBR DX: 496 nm ____Excel. ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ OliGreen: 498 nm ____Excel. ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ RiboGreen: 500 nm ____Excel. ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PicoGreen: 502 nm ____Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Rhodamine Green: 502 nm ____Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ BODIPY FL: 503 nm ____Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Rhodamine 123: 505 nm ____
Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Ethidium bromide: 518 nm ____Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ JOE (6-JOE): 520 nm ____Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ TET: 520 nm ____
Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Rhodamine R6G: 525 nm ____
Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ BODIPY R6G: 528 nm ____Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ HEX: 530 nm ____Med. ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Alexa Fluor 532: 531 nm ____Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ BODIPY 530/550: 535 nm ____Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Ethidium bromide homodimer: 535 nm ____Excel. ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Nile Red: 535 nm ____Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Propidium iodide: 535 nm ____
Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ BODIPY TMR: 542 nm ____Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ TAMRA (5-TAMRA): 542 nm ____Excel. ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ SYPRO Red: 544 nm ____
Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Cy3: 550 nm ____
Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ TM Rhodamine: 554 nm ____
Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Rhodamine Red: 560 nm ____Med. ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Alexa Fluor 546: 561 nm ____
Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ ROX (5-ROX): 567 nm ____
Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Lissamine Rhodamine: 570 nm ____Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ XRITC: 572 nm ____
Med. ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Alexa Fluor 568: 579 nm ____
Med. ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Cy3.5: 581 nm ____
Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ BODIPY TR: 589 nm ____Med. ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Alexa Fluor 594: 590 nm ____
Good ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Texas Red: 596 nm ____
Poor ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Alexa Fluor 633: 631 nm ____
Poor ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Cy5: 650 nm ____
Poor ___ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Cy5.5: 675 nm ____
Emission Maxima
____ 443 nm: Coumarin
____ 461 nm: DAPI
____ 506 nm: Cy2____ 512 nm: BODIPY FL____ 514 nm: Oregon Green____ 517 nm: FAM (5-FAM)____ 518 nm: FITC____ 518 nm: OliGreen____ 519 nm: Alexa Fluor 488____ 520 nm: GelStar____ 520 nm: RiboGreen____ 521 nm: SYBR Green I/SYBR Green II____ 523 nm: PicoGreen____ 526/650 nm: Acridine Orange____ 527 nm: Rhodamine Green
____ 533 nm: Rhodamine 123
____ 537 nm: SYBR Gold____ 540 nm: GFP (Green Fluorescent Protein)____ 540 nm: TET
____ 548 nm: JOE (6-JOE)____ 550 nm: BODIPY R6G____ 550 nm: SYBR DX____ 554 nm: Alexa Fluor 532____ 555 nm: AttoPhos____ 555 nm: BODIPY 530/550____ 555 nm: Rhodamine R6G____ 556 nm: HEX
____ 565 nm: Cy3____ 568 nm: SYPRO Orange____ 568 nm: TAMRA (5-TAMRA)
____ 572 nm: Alexa Fluor 546____ 573 nm: TM Rhodamine____ 574 nm: BODIPY TMR____ 575 nm: Nile Red
____ 578 nm: R-phycoerythrin
____ 580 nm: Rhodamine Red
____ 590 nm: Lissamine Rhodamine____ 590 nm: NanoOrange____ 591 nm: ROX (5-ROX)
____ 595 nm: XRITC____ 596 nm: Cy3.5
____ 603 nm: Alexa Fluor 568____ 605 nm: Ethidium bromide
____ 615 nm: Texas Red____ 617 nm: BODIPY TR____ 617 nm: Ethidium bromide homodimer____ 617 nm: Propidium iodide____ 618 nm: Alexa Fluor 594____ 618 nm: SYPRO Ruby____ 619 nm: SYPRO Red
____ 630 nm: Radiant Red
____ 647 nm: Alexa Fluor 633
____ 670 nm: Cy5
____ 694 nm: Cy5.5
FluorophoreReference GuideFluorophore Excitation and Emission Data
526SP526 nm
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I I I I I I I I
Wavelength (nm) 350 400 450 500 550 600 650 700
For more information on Bio-Rad imaging products,contact your local Bio-Rad representative. In the US,call 1-800-4BIORAD (1-800-424-6723).Visit us on the Web at discover.bio-rad.com
Bio-Rad Laboratories, Inc.Life Science Group2000 Alfred Nobel DriveHercules, CA 94547 USA
Alexa Fluor, BODIPY, NanoOrange, OliGreen, Oregon Green, PicoGreen, RiboGreen, SYBR, SYPRO, and Texas Red aretrademarks of Molecular Probes, Inc. AttoPhos is a trademark of JBL Scientific. Cy is a trademark of Amersham PharmaciaBiotech. FAM, HEX, ROX, and TAMRA are trademarks of Applera Corp. GelStar is a trademark of FMC Corp. Lissamine isa trademark of Imperial Chemical Industries PLC. Nile Red is a trademark of Sigma-Aldrich Co.
Bulletin 2421 Rev C 02-060 0502
Spectra:
Excitation Peak Emission PeakResponse to Broad UV Excitation
ExcitationMaxima
EmissionMaxima
Molecular Imager® FXEmission Filter
VersaDoc™
Imaging System EmissionFilter
FluorophoreReference GuideFluorophore Applications and Spectra
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Inte
nsi
ty
Key:
Fluorophore Name Supplier Application
Alexa Fluor 488 Molecular Probes General conjugated label
Good 495 nm 520 nm 530DF30 530DF60
Alexa Fluor 532 Molecular Probes General conjugated label
Good 531 nm 554 nm 550LP/605DF50 520LP
Alexa Fluor 633 Molecular Probes General conjugated label
Good 631 nm 647 nm 695DF50 Not supported
AttoPhos Molecular Probes General conjugated label
Good 440 nm 555 nm 530DF30 Custom
Coumarin Molecular Probes General conjugated label
Good 402 nm 443 nm Not supported Custom
Cy2 Amersham/Pharmacia General conjugated label
Good 489 nm 506 nm 530DF30 530DF60
Cy3 Amersham/Pharmacia General conjugated label
Good 550 nm 565 nm 555LP 520LP
Cy3.5 Amersham/Pharmacia General conjugated label
Medium 581 nm 596 nm 555LP 520LP
Cy5 Amersham/Pharmacia General conjugated label
Good 650 nm 670 nm 695DF55 Not Supported
Ethidium bromide Bio-Rad DNA stain
Good 518 nm 605 nm 555LP 520LP
FITC/FAM Molecular Probes General conjugated label
Good 494 nm 518 nm 530DF30 530DF60
eGFP (Green Fluorescent Protein) Clontech Reporter gene assay
Poor 488 nm 509 nm 530DF30 520LP
HEX ABI General conjugated label
Good 530 nm 556 nm 555LP 520LP
JOE (6-JOE) ABI General conjugated label
Good 520 nm 548 nm 530DF30 520LP
Lissamine Rhodamine Molecular Probes General conjugated label
Good 570 nm 590 nm 555LP 520LP
NanoOrange Molecular Probes Proteins in solution
Excellent 485 nm 590 nm 555LP 520LP
Nile Red Sigma Protein gel stain
Excellent 535 nm 575 nm 555LP 520LP
OliGreen Molecular Probes ssDNA and oligos in solution
Excellent 498 nm 518 nm 530DF30 520LP
Oregon Green Molecular Probes General conjugated label
Excellent 490 nm 514 nm 530DF30 520LP
PicoGreen Molecular Probesd sDNA in solution
Excellent 502 nm 523 nm 530DF30 520LP
Propidium iodide Molecular Probes DNA and cell stain
Good 535 nm 617 nm 555LP/640DF35 610LP
Radiant Red Bio-Rad RNA gel stain
Excellent 495 nm 630 nm 555LP/640DF35 610LP
Rhodamine 123 Molecular Probes General conjugated label
Good 505 nm 533 nm 530DF30 520LP
RiboGreen Molecular Probes RNA in solution
Excellent 500 nm 520 nm 530DF30 520LP
ROX (5-ROX) ABI General conjugated label
Good 567 nm 591 nm 555LP 520LP
R-phycoerythrin Molecular Probes General conjugated label
Good 480/565 nm 578 nm 555LP/530DF30 520LP
SYBR Gold Molecular Probes dsDNA and RNA gel stain
Excellent 495 nm 537 nm 530DF30 520LP
SYBR Green I Molecular Probes dsDNA and RNA gel stain
Excellent 494 nm 521 nm 530DF30 520LP
SYBR Green II Molecular Probes ssDNA and RNA gel stain
Excellent 492 nm 521 nm 530DF30 520LP
SYPRO Orange Bio-Rad Protein gel stain
Excellent 480 nm 568 nm 555LP/530DF30 520LP
SYPRO Red Molecular Probes Protein gel stain
Excellent 544 nm 619 nm 555LP/640DF35 520LP
SYPRO Ruby Molecular Probes Protein gel stain
Excellent 300/480 nm 618 nm 555LP 610LP
TAMRA (5-TAMRA) ABI General conjugated label
Good 542 nm 568 nm 555LP 520LP
Texas Red Molecular Probes General conjugated label
Good 596 nm 615 nm 640DF35 610LP
XRITC Molecular Probes General conjugated label
Good 572 nm 595 nm 555LP/640DF35 520LP