常见荧光染料及波长选择.xls

Page 1 Rev 8 29 05 Fluorescence Indicators and Filter Wavelength List FLUORESCENT DYE KITCOMPANYEXCITEEMITPOSSIBLE FILTERSAPPLICATION 7 AAD546647540 35 645 40Nucleic Acid Stain 545 40 645 40 540 25 645 40 Acridine OrangeSigma A6014490519485 20 528 20Nucleic Acid Stain 485 20 530 25Other peaks include 502 526 Acridine Orange460650460 40 645 40Bound to RNA Acridine YellowSigma A2520455515450 50 516 20 450 50 528 20 450 50 530 25 Acriflavine HCISigma A8251465510460 40 530 25 460 40 516 20 Acriflavine NeutralSigma A8126463 261517460 40 530 25 Allophycocyanine620 40 680 30General Fluorescent Tag Alamar BlueBiosource560590540 35 590 20Cell Viability Dye 540 25 590 20Better than MTT XTT 540 25 590 35Thymidine or Neutral 560 15 590 20Red Nontoxic to cells 565 10 590 10Also can be read at 570 530 25 590 35600 in absorbance mode 528 20 590 35 600 40 can also be used for emission Alexa Fluor 350 DAPI Blue 347442340 30 440 40Other citations for peaks 340 30 450 50include 346 for ex 340 30 460 40 360 40 460 40 Alexa Fluor 430Molecular Probes430550420 50 560 40Other citations for peaks Page 2 420 50 560 20include 431 541 420 50 545 40 Alexa Fluor 488Molecular Probes493517485 20 528 20Other citations for peaks 485 20 530 25include 495 519 Alexa Fluor 532Molecular Probes532554530 25 565 10 528 20 560 20FLx800 only 516 20 560 20 Alexa Fluor 546Molecular Probes556573540 25 590 20 540 25 590 35 540 25 600 40 540 35 600 40 Alexa Fluor 555Molecular Probes555565540 25 590 20Similar to tetramethyl 530 25 590 20rhodamine and CY3 528 20 590 20More photostable 540 25 590 35conjugates brighter 540 25 600 40 530 25 590 35Filters listed in order of signal noise Alexa Fluor 568Molecular Probes577603560 20 620 40575 15 620 40 might be able to be used but would require cheating Alexa Fluor 594Molecular Probes588613590 20 620 15Other citations for peaks 590 10 635 32include 590 617 590 20 635 32 Alexa Fluor 633Molecular Probes632647620 15 645 15 Alexa Fluor 647Molecular Probes650668620 40 680 30Best combination filt set 645 15 680 30Cheating is required 635 32 680 30Cheating is required 635 10 680 30 Alexa Fluor 660Molecular Probes663690655 20 702 20Filter set does not exist yet Alexa Fluor 680Molecular Probes679702670 20 702 20Filter set does not exist 684707670 20 702 20yet Useful in multicolor app with TMR Texas Red and R phycoerythrin Alexa Fluor 700Molecular Probes696719680 30 730 20Filter set does not exist yet Page 3 Alexa Fluor 750Molecular Probes752775745 20 785 20Filter set does not exist yet Allophycocyanin650660620 40 680 30Not tried AlphascreenPerkin Elmer680520 620680 30 570 100Requires Synergy instrument and 2 filter sets o aminobenzoyl310410310 20 420 50 310 20 410 10 310 20 400 30 2 aminobenzoic acid320420320 20 420 50 340 30 420 50 9 Aminoacridine HCISigma A1135423458400 30 460 40 Aminocoumarin350445340 30 460 40 340 30 450 50 360 40 460 40 Aminomethylcoumarin AMCA KPL353440360 40 460 40Other sources indicate 340 30 460 40350 450 as peaks 340 30 440 40 6 aminoquinoline 6 AQ 355550360 40 560 40 7 amino trifluoromethyl coumarin400505400 30 508 20Caspace Substrate AFC 400 10 508 20 516 20 could also be em 7 amino 4 methylcoumarin 7 AMC 351430340 30 440 40Caspace Substrate 340 30 440 30 360 40 440 40 360 40 440 30 Anilino 1 naphthalene sulfonic acidSigma A1028385485380 20 485 40 380 20 485 20 Amplex RedMolecular Probes563587540 35 590 20Any combination of the A 122 10540 35 600 40filters to the left should 540 25 590 35work fine 540 25 600 40 560 15 590 20 540 25 590 20 565 10 590 10 530 25 590 35 Page 4 530 25 600 40 Attophos TM JBL Scientific420560440 30 560 40Immunoassay substrate 450 50 580 50Other sources indicate 435 555 as peaks Atto Tag CBQCA456560450 50 560 40 BBOT 2 5 bis 5 tert Butylbenz Sigma B7878372433360 40 440 30 oxazolyl 2 Thiophene 360 40 440 40 BCECF500530485 20 530 25Ester as Vital Cell Dye 485 20 540 25pH Indicator Other citations include 501 527 Ben Zoxanthene Yellow360 40 460 40General Fluorescent Tag 7 Benzylamino 4 BBD Sigma B7764315377310 20 380 20 4 Biphenylyl PBBO Sigma B3386328391 411340 30 400 30 Bis ANSMolecular Probes395500400 30 500 27Best filter set B153380 20 508 20 Bodipy 581 591Molecular Probes581591575 15 620 40Cheating required D3861575 15 620 15Might be the best filter set 560 40 620 40Cheating required 560 40 620 15 560 20 620 40 560 15 590 20 Bodipy FLMolecular Probes505513485 20 530 25General Fluorescent Tag 485 20 528 20Used in Enchek Kit Other citations include 502 511 as peaks Bodipy TMRMolecular Probes542574530 25 590 35 Bodipy TRMolecular Probes589617590 20 635 32 590 20 645 40 590 10 635 32 Bodipy 650Molecular Probes647661620 40 680 30 635 32 680 30Some cheating required 645 15 680 30Some cheating required BrdU Cell Proliferation Assay HTSCalbiochem325420320 20 420 50Other ex filters include 310 20 340 11 340 30 Page 5 Other em filters include 370 470 nm filters BRET2PE390 400505 510410 80 and 515 30 for dualThese are filters recommended by PE emission 2 4 t Butylphenyl 5 4 Biphenylyl Sigma B8378307362 367310 20 360 40 1 3 4 Oxadiazole Butyl PBD Calcein AMMolecular Probes493517485 20 528 20Viability Assays 485 20 530 25 Calcium Green 1Molecular Probes506531485 20 530 25Calcium Uptake 485 20 528 20 485 20 540 25 500 27 540 25 Calcium Green 2Molecular Probes503 506536485 20 530 25Calcium Uptake 500 27 540 25 Calcium OrangeMolecular Probes549575 576540 25 590 35Calcium Uptake Calcium CrimsonMolecular Probes589 590615575 15 635 32 590 20 635 32 590 10 635 32 Carbostyril 124Sigma C9027351413360 40 440 40 340 30 400 30 340 30 420 50 Carboxy FluoresceinSigma C7153494518485 20 530 25Vital Dye pH Indicator 485 20 528 20Liposome cell interations Carboxyfluorescein diacetateMolecular Probes492515485 20 530 25 C 1361485 20 516 20 485 20 528 20 Caspace 3Assay Designs360440360 40 440 40Best filter combo 360 40 440 30AMC is the substrate 360 40 460 40 Caspace 3Calbiochem365 380430 460360 40 440 40 360 40 440 30 360 40 450 50 360 40 460 40 Caspace 8R D Systems400505400 30 508 20 Page 6 CBQCAMolecular Probes465560450 50 560 40Protein quantitation A 6222450 50 580 50 460 40 545 40 CellQuanti BlueBioAssay Systems530 570590 620540 35 600 40Cell Proliferation and Cytotoxicity 530 25 590 35 540 25 590 20 Cell Tracker OrangeMolecular Probes541565528 20 575 15Not tried C 2927530 25 575 15 530 25 590 35 Cell Titer BluePromega579584560 15 590 20Listed filters not tried G8080565 10 590 10If available 540 35 590 20 540 35 600 40This pair works well 540 25 590 35 540 25 600 40 530 25 600 40 CFDA498523485 20 530 25Extracellular pH Peaks guessed at from spectra CFDA AM485 20 530 25Plasma Membrane Disruption Chlorophyll444683440 40 680 30 440 30 680 30 420 50 680 30 Cis parinaric acid320416320 20 420 50 CM Dil553570540 25 590 20Listed in order of signal noise 530 25 590 20based on the performance of these 528 20 590 20filter pairs with Cy3 a compound with 540 25 590 35similar spectra 540 25 600 40 530 25 590 35 Coproporphyrin405610400 30 620 40 395 25 620 40 Coumarin 4 4 MU MUB Sigma M1381360454360 40 460 40 Coumarin 35Sigma C9152401489400 30 485 40 Coumarin 120Sigma A9891354440360 40 440 40 Coumarin 138C9902365460360 40 460 40 Page 7 Coumarin 151Sigma A8401382484360 40 485 40 380 20 485 40 Coumarin 152Sigma C9527397519395 25 530 25 400 30 530 25 Coumarin 153Sigma C9777423538420 50 530 25 420 50 540 25 Coumarin 311Sigma C9402368453360 40 460 40 CFP 1440480440 30 485 20 420 50 485 20 440 40 500 27 Cresyl Violet CIO4Sigma C0664503615500 27 620 40 CUG386448380 20 460 40Preferred 380 20 450 50 380 20 440 30 380 20 440 40 360 40 460 40 360 40 450 50 360 40 440 40 360 40 440 30 Cy2KPL489505485 20 530 25Other sources indicate 485 20 528 20490 508 as peaks 485 20 516 20 Jackson Immuno489510same as aboveMore photostable than FITC Less sensitive to pH More fluorescent in organic mounting media Cy3Biol Detect Sys550565540 25 590 20Protein and modified Res Organics552568530 25 590 20oligonucleotide labelling 528 20 590 20 540 25 590 35Listed in order of signal to noise 540 25 600 40 530 25 590 35 Amersham550570same as above Cy3 5Amersham581596575 15 620 40 Page 8 Cy5Biol Detect Sys652667620 40 680 30Protein and modified Res Organics650670635 32 680 30oligonucleotide labelling 635 10 680 30620 40 is best for ex Cheating required Amersham649 650670620 40 680 30 635 32 680 30 Cy5 5Amersham675694665 20 702 20Filter set does not exist yet Cy7Amersham743767730 20 770 20These are theoretical wavelengths not available yet CyQuantMolecular Probes502523485 20 528 20Both sets are equal 485 20 530 25 500 27 540 25 CyQuant NFMolecular Probes502523485 20 528 20Streamlined version of C35007485 20 530 25Cyquant 500 27 540 25 CytoTox OnePromega560590540 35 590 20 G7890 7892540 35 600 40 540 25 590 35 540 25 600 40 540 25 590 20 560 15 590 20 DABA400 30 508 20 DAR 4MALX 620 067560575540 35 600 40 540 25 590 35 560 15 590 20 540 25 590 20 Dansyl ChlorideSigma D2625355500360 40 500 27Reacts with proteins to form fluorescent peptide derivatives Dansyl P E 360 40 530 25General Fluorescent Tag DDAO648657590 20 645 40Very broad ex peak 540 648633 700575 15 645 40680 30 can also be used for emission 560 40 645 40540 35 can also be used for excitation 560 20 645 40 Diethylthiacarbo cyanine iodideSigma D2409662679635 32 680 30 Page 9 Diaminofluorescein diacetate492515485 20 530 25 485 20 528 20 6 Diamond 2 Phenylindole DAPI Sigma D9542372456360 40 460 40 359461360 40 460 40 5 Dichloro 11 diphenylamino 3Sigma I8258823950None recommended IR 140 DibromofluoresceinSigma D3017505568500 27 560 40 DichlorofluoresceinSigma D6132517544 2508 20 560 40 Dichlorofluorescein495518485 20 530 25 485 20 528 20 Dihydroethidium518605516 20 620 40Oxidized version 508 20 620 40600 40 can also be used 500 27 620 40for emission 528 20 620 40 Dil549 550565540 25 590 20 530 25 590 20 528 20 590 20 540 25 590 35 540 25 600 40 530 25 590 35 2 Diphenyl 1 PPD Sigma D5900281341281 10 340 30Synergy HT TR only 4 Diphenyl 1 DPB Sigma D5268315 345430360 40 450 50 340 30 450 50 6 Diphenyl 1 DPH Sigma D4380351430360 40 450 50 340 30 450 50 dipyridamole450520450 50 528 20 433500440 40 528 20 440 30 528 20 dipyridamole433500420 50 500 27 Dnase Alert kitAmbion 1970490520485 20 528 20 Doxorubicin488510485 20 516 20 485 20 528 20Might be the best filter 485 20 530 25set Page 10 DTAF485 20 530 25Alcohol Reactive Probe Dylight 547Pierce557574540 25 590 20Brighter than Cy3 530 25 590 20590 35 is better 528 20 590 20 540 25 590 35 540 25 600 40 530 25 590 35 Dylight 647Pierce652673620 40 680 30Brighter than Cy5 635 32 680 30620 40 is better Dylight 680Pierce682715Available from Omega Dylight 800Pierce770794Available from Omega EDANS335493340 30 500 27 340 30 485 20 340 30 485 40 320 20 as above ELF 97 alcohol345530340 30 530 25 340 30 540 35 EnzChek Amylase Assay KitMolecular Probes505512485 20 528 20Bodipy is the fluorophore E 11954485 20 530 25Kinetic assay 485 20 516 20 EnzChek MPO activity assay kitMolecular Probes485530485 20 528 20Testing aminophenyl E33856485 20 530 25fluorescein 530590530 25 590 35Testing amplex red see amplex red for other choices EnzChek Protease Assay Kit GreenMolecular Probes505513485 20 528 20 E 6638485 20 530 25 485 20 516 20 EnzChek Protease Assay Kit RedMolecular Probes589617590 20 645 40 E 6639590 10 635 32 590 10 645 40 EnzoLyte 490 HIV Protease Anaspec340490340 30 500 27 Assay Kit Fluorimetric71127340 30 485 40 340 11 485 20 EnzoLyte 520 HIV ProteaseAnaspec490520485 20 528 20 Assay Kit Fluorimetric71147 Page 11 EnzoLyte 490 HCV ProteaseAnaspec340490340 30 500 27 Assay Kit Fluorimetric71126340 30 485 40 340 11 485 20 EnzoLyte 520 HCV ProteaseAnaspec490520485 20 528 20 Assay Kit Fluorimetric71145 EnzoLyte 620 HCV ProteaseAnaspec590620590 20 620 15 Assay Kit Fluorimetric71146590 20 645 40 590 20 635 32 Eosin YSigma B6003522551508 20 560 40 516 20 580 50 EpicoccononeFluorotechnics400610400 30 620 40 480 540610485 540 620 40 Ethidium Bromide Sigma E8751545610540 35 620 40Dye Bound to DNA 530 25 620 40 540 25 620 40 518605516 20 600 40 516 20 620 40 Ethidium Bromide in sol n 485 40 620 40Nucleic Acid Stain Ethidium Homodimer485 40 590 35Nucleic Acid Stain 528617528 20 620 40 528 20 620 15 530 25 as above Ethoxyresorufin544590540 25 600 40Other sources indicate 540 25 590 20535 585 peaks 540 25 590 10 530 25 590 35 Europium340615360 620 40Requires Synergy HT with TRF 360 ex using monochromator works better than 340 ex Evans BlueSigma E2129550610540 25 620 40 FAM492518485 20 530 25 485 20 528 20 Fluo 3503526485 20 530 25Ca2 Concentration 485 20 528 20 Page 12 506527500 27 540 25 Fluo 4494516485 20 530 25 485 20 528 20 Fluorescamine390480380 20 485 40Protein quantitation Molecular Probes380464380 20 460 40 F 2332395 25 485 40 400 30 460 40 380 20 485 20 360 40 460 40 Fluorescein 5 isothiocyanate FITC Sigma F7250422546420 50 545 40 485 20 530 25 Fluoroblok tests485 20 528 20For Calcein AM Often use Calcein AM and Dil540 25 590 20For Dil FITCKPL495528485 20 530 25Other sources indicate 485 20 528 20492 520 as peaks Fluorescein DiacetateSigma F7378490541 571485 20 560 40 485 20 528 20 485 20 530 25 Fluorescein SodiumSigma F6377490550485 20 530 25Monolayer Permeability 485 20 528 20 Fluorescein490513485 20 528 20 485 20 530 25 FluorXBiol Detect Sys494520485 20 528 20Protein antibodies syn Res Organics485 20 530 25thetic oligonucleotide Fura 2Molecular Probes338 366505340 30 380 20 for exCalcium Uptake 508 20 for em 340 11 380 20 for ex 508 20 for em GFP BFP RFP variants ASredClontech576592560 15 590 20 EBFP380460380 20 460 40 380440380 20 440 40 380 20 440 30 380 20 450 50 360 40 460 40 Page 13 EGFP489509485 20 516 20 485 20 530 25 485 20 528 20 pECFP433475420 50 485 20This combo works well according to user GFPuv395509400 30 508 20 380 20 508 20 360 40 508 20 GFP S65T489509485 20 530 25 HcRedClontech588618590 20 635 32First pair should be best 590 10 635 32645 40 might also work 575 15 635 32ok as an emission filter 560 40 635 32 RFPClontech558583540 35 600 40First pair should be best 540 25 590 20 540 25 590 35 Stemmer395509400 30 508 20 380 20 508 20 360 40 508 20 wtGFP395509395 25 508 20Minor peak at 470nm 400 30 508 20Compare 395 25 508 20 380 20 508 20with 485 20 516 20 360 40 508 20528 20 and 530 25 470509485 20 516 20Minor ex peak 489508485 20 528 20 485 20 530 25 460 40 516 20 Y66H382459360 40 460 40 pEYFPClontech514527500 27 540 25Best 485 20 528 20 485 20 530 25 500 27 560 15Using this set resulted in 4 5 1 s n ratio for one customer Yellow Fluorescent Protein525535 540500 27 540 25 516 20 560 40 Page 14 H2DCF488515485 20 528 20 HEXEpoch535556530 25 565 10Listed in order of 528 20 565 10performance of 530 25 590 35HEX FAM ratio 528 20 590 35 530 25 575 15 528 20 575 15 HIDC IodideSigma H7636637740None recommended HITC IodideSigma H0387740879None recommended HNPPRoche 553584540 25 590 20 11578888001530 25 590 20 528 20 590 20 540 25 590 35 540 35 600 40 Hoechst 33258Sigma B2883365465360 40 460 40DNA dye 345478340 30 460 40 340 30 485 40 Hoechst 33342Sigma B2261355468360 40 460 40DNA dye 343483340 30 460 40 340 30 485 40 7 hydroxycoumarin380 20 460 40 360 40 460 40 Hydroxymethylcoumarine360 40 460 40Immunoassay 3 p Hydroxyphenyl proprionic acid320405320 20 400 30Best filter set by far otherwise known as HPPA310 20 400 30Not nearly as good as 320 400 filter set p Hydroxy phenylacetic 4 HPA 315425310 20 420 50 320 29 420 50 Indo 1Sigma I3136355405 485360 40 for exCalcium Ion Concen 420 50 485 20 for em 340 30 for ex 400 30 485 40 for em Indocyanine green780 800810 830None available IQ kinase assayPierce Chemical560590560 15 590 20 560 15 590 10 Page 15 Isoniaside396465395 25 460 40 395 25 485 40 JC1Molec Probes514529500 27 540 25Mitochondrial detection T3168475 485530 590485 20 530 25 485 20 540 35 485 20 540 25 590 35 or 600 40Ratioed emission filters KDalert GAPDHAmbion 1639560590540 35 600 40 540 35 590 20 560 15 590 20 540 25 590 35 Laurodan360 40 460 40Membrane Probe LIVE DEAD Cell vitality assay kitMolec Probes488575485 20 575 15 L34951485 20 580 50 485 20 590 35 Lucifer YellowSigma L0259 440 30 530 25Gap Junc n Comm n L0144Marks nerve cells 425530420 50 528 20 420 50 530 25 420 50 540 25 420 50 540 35 428540420 50 540 35 Lysosensor Yellow Blue low pH384540380 20 540 35 380 20 540 25 360 40 540 35 7 methoxycoumarin328393340 30 400 30Fluorogenic substrate for matrix metalloproteinases Mca Ala7 Lys Dnp 9 BradykininBachem M 2405328393340 30 400 30Fluorogenic substrate for 340 30 395 25matrix metalloproteinases MDA508 20 560 40 Measure iT Nitrate Assay kitMolecular Probes365450360 40 460 40 Measure iT Thiol Assay kitMolecular Probes494517485 20 528 20 Measure iT Lead Assay kitMolecular Probes495516485 20 528 20 Page 16 Measure iT Cadmium Assay kitMolecular Probes495516485 20 528 20 Methylumbelliferyl beta D 316376310 20 380 20 galactopyranoside MUG 4 methylumbelliferyl phospate370430360 40 440 30 4 MUP 360 40 440 40 Methylumbelliferone360 40 460 40Immunoassay Substrate 4 MeU 4 methylumbelliferone 360448360 40 460 40 7 hydroxy 4 methylcoumarin 360 40 450 50 360 40 440 40 Mithramycin450570450 50 580 50 440 40 590 35 440 40 575 15 440 30 580 50 Mito PT488590485 40 590 35 485 20 590 35 Mithramycin ASigma M1381401556400 30 560 40 400 10 560 40 MitoTracker Green FMMolecular Probes490516485 20 528 20Mitochondria Stain M 7514485 20 530 25 485 20 530 25 MitoTracker Orange CMTMRosMolecular Probes554576540 25 590 20Mitochondria Stain M 7510530 25 590 20Listed in order of signal to noise 528 20 590 20 540 25 590 35 540 25 600 40 530 25 590 35 MitoTracker Orange CM H2TMRosMolecular Probes554576540 25 590 20Mitochondria Stain M 7511530 25 590 20 528 20 590 20 540 25 590 35 540 25 600 40 530 25 590 35 MitoTracker Red CMXRosMolecular Probes579599560 20 620 40Mitochondria Stain M 7512575 15 620 40Have to enter 620 40 as 620 35 MitoTracker Red CM H2XRosMolecular Probes579599560 20 620 40Mitochondria Stain Page 17 M 7513575 15 620 40Have to enter 620 40 as 620 35 Monobromo380475380 20 485 40 380 20 485 20 Monobromobimane390