Fluorophore Reference Guide Fluorophore Excitation and Emission Data

Laser Lines Broad UV Excitation Excitation Maxima Emission Maxima Emission Filters 290-365 nm LP = Long pass filter DF = Band pass filter Excel. ______DAPI: 359 nm ____ SP = Short pass filter Good ______GFP (Green Fluorescent Protein): 395 nm ____ 400 nm Good ______Coumarin: 402 nm ____

425 nm

Good ______AttoPhos: 440 nm ____

____ 443 nm: Coumarin

450 nm

Good ______Acridine Orange: 460/500 nm ______461 nm: DAPI

Good ______R-phycoerythrin: 480/565 nm ____ Excel. ______SYPRO Orange/SYPRO Ruby: 480 nm ____ 475 nm Excel. ______NanoOrange: 485 nm ____ Good ______Cy2: 489 nm ____ ArI 488 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 ____ Molecular Imager FX Filters VersaDoc Filters Excel. ______SYBR Gold: 495 nm ____ Good ______SYBR DX: 496 nm ____ Excel. ______OliGreen: 498 nm ____ 500 nm Excel. ______RiboGreen: 500 nm ____ 500 nm Excel. ______PicoGreen: 502 nm ____ Good ______Rhodamine Green: 502 nm ____ Good ______BODIPY FL: 503 nm ____ Good ______Rhodamine 123: 505 nm ____ ArI 514 nm ____ 506 nm: Cy2 ____ 512 nm: BODIPY FL ____ 514 nm: Oregon Green 515 nm ____ 517 nm: FAM (5-FAM) ____ 518 nm: FITC Good ______Ethidium bromide: 518 nm ______518 nm: OliGreen 520 nm Good ______JOE (6-JOE): 520 nm ______519 nm: Alexa Fluor 488 520LP Good ______TET: 520 nm ______520 nm: GelStar ____ 520 nm: RiboGreen ____ 521 nm: SYBR Green I/SYBR Green II Good ______Rhodamine R6G: 525 nm ____ 525 nm ____ 523 nm: PicoGreen ____ 526/650 nm: Acridine Orange 526SP Good ______BODIPY R6G: 528 nm ______527 nm: Rhodamine Green 526 nm DPSSL Good ______HEX: 530 nm ____ 530DF30 530DF60 (µGreen) 532 nm Med. ______Alexa Fluor 532: 531 nm ____ Good ______BODIPY 530/550: 535 nm ______533 nm: Rhodamine 123 Good ______Ethidium bromide homodimer: 535 nm ____ Excel. ______Nile Red: 535 nm ______537 nm: SYBR Gold Good ______Propidium iodide: 535 nm ______540 nm: GFP (Green Fluorescent Protein) ____ 540 nm: TET Good ______BODIPY TMR: 542 nm ____ Good ______TAMRA (5-TAMRA): 542 nm ____ GHeNe 543 nm Excel. ______SYPRO Red: 544 nm ______548 nm: JOE (6-JOE) 545 nm Good ______Cy3: 550 nm ____ 550 nm ____ 550 nm: BODIPY R6G ____ 550 nm: SYBR DX Good ______TM Rhodamine: 554 nm ______554 nm: Alexa Fluor 532 ____ 555 nm: AttoPhos 555 nm ____ 555 nm: BODIPY 530/550 555LP ____ 555 nm: Rhodamine R6G ____ 556 nm: HEX Good ______Rhodamine Red: 560 nm ____ 560 nm Med. ______Alexa Fluor 546: 561 nm ______565 nm: Cy3 Good ______ROX (5-ROX): 567 nm ______568 nm: SYPRO Orange ____ 568 nm: TAMRA (5-TAMRA) Good ______Lissamine Rhodamine: 570 nm ____ Good ______XRITC: 572 nm ______572 nm: Alexa Fluor 546 ____ 573 nm: TM Rhodamine ____ 574 nm: BODIPY TMR 575 nm ____ 575 nm: Nile Red

____ 578 nm: R-phycoerythrin Med. ______Alexa Fluor 568: 579 nm ____ 580 nm ____ 580 nm: Rhodamine Red Med. ______Cy3.5: 581 nm ____

____ 590 nm: Lissamine Rhodamine Good ______BODIPY TR: 589 nm ______590 nm: NanoOrange Med. ______Alexa Fluor 594: 590 nm ______591 nm: ROX (5-ROX) YHeNe 594 nm ____ 595 nm: XRITC Good ______Texas Red: 596 nm ______596 nm: Cy3.5 605DF50 600 nm ____ 603 nm: Alexa Fluor 568 ____ 605 nm: Ethidium bromide 610 nm 610LP ____ 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 622.5 nm 625 nm

630 nm ____ 630 nm: Radiant Red Poor ______Alexa Fluor 633: 631 nm ____ Red diode 635 nm

640DF35

____ 647 nm: Alexa Fluor 633

Poor ______Cy5: 650 nm ____ 650 nm

657.5 nm 660 nm 660 nm ____ 670 nm: Cy5 670 nm Poor ______Cy5.5: 675 nm ____ 690DF50 ____ 694 nm: Cy5.5 Fluorophore Reference Guide Fluorophore Applications and Spectra

Key: Spectra:

Fluorophore Name Supplier Application

VersaDoc™ ® Excitation Peak Emission Peak Response to Broad Excitation Emission Molecular Imager FX Imaging System Emission UV Excitation Maxima Maxima Emission Filter Filter Intensity

100 Alexa Fluor 488 Molecular Probes General conjugated label 80 60 40 Good 495 nm 520 nm 530DF30 530DF60 20

100 Alexa Fluor 532 Molecular Probes General conjugated label 80 60 40 Good 531 nm 554 nm 550LP/605DF50 520LP 20

100 Alexa Fluor 633 Molecular Probes General conjugated label 80 60 40 Good 631 nm 647 nm 695DF50 Not supported 20

100 AttoPhos Molecular Probes General conjugated label 80 60 40 Good 440 nm 555 nm 530DF30 Custom 20

100 Coumarin Molecular Probes General conjugated label 80 60 40 Good 402 nm 443 nm Not supported Custom 20

100 Cy2 Amersham/Pharmacia General conjugated label 80 60 40 Good 489 nm 506 nm 530DF30 530DF60 20

100 Cy3 Amersham/Pharmacia General conjugated label 80 60 40 Good 550 nm 565 nm 555LP 520LP 20

100 Cy3.5 Amersham/Pharmacia General conjugated label 80 60 40 Medium 581 nm 596 nm 555LP 520LP 20

100 Cy5 Amersham/Pharmacia General conjugated label 80 60 40 Good 650 nm 670 nm 695DF55 Not Supported 20

100 Ethidium bromide Bio-Rad DNA stain 80 60 40 Good 518 nm 605 nm 555LP 520LP 20

100 FITC/FAM Molecular Probes General conjugated label 80 60 40 Good 494 nm 518 nm 530DF30 530DF60 20

100 eGFP (Green Fluorescent Protein) Clontech Reporter gene assay 80 60 40 Poor 488 nm 509 nm 530DF30 520LP 20

100 HEX ABI General conjugated label 80 60 40 Good 530 nm 556 nm 555LP 520LP 20

100 JOE (6-JOE) ABI General conjugated label 80 60 40 Good 520 nm 548 nm 530DF30 520LP 20

100 Lissamine Rhodamine Molecular Probes General conjugated label 80 60 40 Good 570 nm 590 nm 555LP 520LP 20

100 NanoOrange Molecular Probes Proteins in solution 80 60 40 Excellent 485 nm 590 nm 555LP 520LP 20

100 Nile Red Sigma Protein gel stain 80 60 40 Excellent 535 nm 575 nm 555LP 520LP 20

100 OliGreen Molecular Probes ssDNA and oligos in solution 80 60 40 Excellent 498 nm 518 nm 530DF30 520LP 20

100 Oregon Green Molecular Probes General conjugated label 80 60 40 Excellent 490 nm 514 nm 530DF30 520LP 20

100 PicoGreen Molecular Probesd sDNA in solution 80 60 40 Excellent 502 nm 523 nm 530DF30 520LP 20

100 Propidium iodide Molecular Probes DNA and cell stain 80 60 40 Good 535 nm 617 nm 555LP/640DF35 610LP 20

100 Radiant Red Bio-Rad RNA gel stain 80 60 40 Excellent 495 nm 630 nm 555LP/640DF35 610LP 20

100 Rhodamine 123 Molecular Probes General conjugated label 80 60 40 Good 505 nm 533 nm 530DF30 520LP 20

100 RiboGreen Molecular Probes RNA in solution 80 60 40 Excellent 500 nm 520 nm 530DF30 520LP 20

100 ROX (5-ROX) ABI General conjugated label 80 60 40 Good 567 nm 591 nm 555LP 520LP 20

100 R-phycoerythrin Molecular Probes General conjugated label 80 60 40 Good 480/565 nm 578 nm 555LP/530DF30 520LP 20

100 SYBR Gold Molecular Probes dsDNA and RNA gel stain 80 60 40 Excellent 495 nm 537 nm 530DF30 520LP 20

100 SYBR Green I Molecular Probes dsDNA and RNA gel stain 80 60 40 Excellent 494 nm 521 nm 530DF30 520LP 20

100 SYBR Green II Molecular Probes ssDNA and RNA gel stain 80 60 40 Excellent 492 nm 521 nm 530DF30 520LP 20

100 SYPRO Orange Bio-Rad Protein gel stain 80 60 40 Excellent 480 nm 568 nm 555LP/530DF30 520LP 20

100 SYPRO Red Molecular Probes Protein gel stain 80 60 40 Excellent 544 nm 619 nm 555LP/640DF35 520LP 20

100 SYPRO Ruby Molecular Probes Protein gel stain 80 60 40 Excellent 300/480 nm 618 nm 555LP 610LP 20

100 TAMRA (5-TAMRA) ABI General conjugated label 80 60 40 Good 542 nm 568 nm 555LP 520LP 20

100 Texas Red Molecular Probes General conjugated label 80 60 40 Good 596 nm 615 nm 640DF35 610LP 20

100 XRITC Molecular Probes General conjugated label 80 60 40 Good 572 nm 595 nm 555LP/640DF35 520LP 20 IIIIIIII Wavelength (nm) 350 400 450 500 550 600 650 700

Alexa Fluor, BODIPY, NanoOrange, OliGreen, Oregon Green, PicoGreen, RiboGreen, SYBR, SYPRO, and Texas Red are For more information on Bio-Rad imaging products, Bio-Rad Laboratories, Inc. trademarks of Molecular Probes, Inc. AttoPhos is a trademark of JBL Scientific. Cy is a trademark of Amersham Pharmacia contact your local Bio-Rad representative. In the US, Life Science Group Biotech. FAM, HEX, ROX, and TAMRA are trademarks of Applera Corp. GelStar is a trademark of FMC Corp. Lissamine is call 1-800-4BIORAD (1-800-424-6723). 2000 Alfred Nobel Drive a trademark of Imperial Chemical Industries PLC. Nile Red is a trademark of Sigma-Aldrich Co. Visit us on the Web at discover.bio-rad.com Hercules, CA 94547 USA

Bulletin 2421 Rev C 02-060 0502