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Emission Filter Guide for Various Fluorescent Reagent
(Excitation and Emission wavelengths)


Fluorescence Reagent ・ Ex/Em wavelength ・ LED light ・ Filterwork List
Below are one of the most popular reagents we often get inquiry.
Click
Spectral button to jump to filterwork guide for each reagent.
Reagent EX nm EX
(Sub)
EM nm spectral
info
Reagent EX nm EX
(Sub)
EM nm spectral
info
Alexa Fluor 532 532 554 GFP 488 507
Cy3 550 570 mCherry 580 610
DsRed 557 579 SYBR Gold 495 540
EtBr 300 518 605 SYBR Green 498 522
FITC 490 525 SYPRO Red 550 300 630
Gel Green 250 500 530 SYPRO Ruby 280 450 620
Gel Red 270 510 600 TagRFP 555 583

 ** Please refer Reagent List page for Ex/Em info of other reagent ⇒  

Notice : The guide is reference purpose only. It does not gurantee any visibility of certain emission.
       Result may vary depends on such factors as LED brightness, filter quality, and camera spec.
 
* ⇒ Long cut filter may be required to eliminate background
■ AlexaFluor532R

★ Place pointer on above figure to see LED & Filter wavelength
For labeling the cellular tissue under the fluorescence microscope and for other studies of cell biology. It holds sulfonic acid which is directly connected to the fluorescent group. Especially bright with rhodamine derivative. The reagent with great light stability

 Life Technology

Em/Ex are close to each other and difficult to separate absorption and excitation spectra. In this case, 505nm and SC-56 combination is effective.
When transmitted light is used, filter to cut off long wavelength side is required for eliminating background.
 Ex  Effcy filter evaluation
 SC-56  SC-60
 470nm      
 505nm      *  
 530nm      
■ Cy3R

★ Place pointer on above figure to see LED & Filter wavelength
In addition to such biological assays as immunocytochemistry, Cyanine dye fluorescence reagent is often used for labelling RNA.

Mirus Bio LLC

Similar to AlexaFluor532, Em/Ex are close to each other and difficult to separate absorption and excitation spectra. In this case, 505nm and SC-56 combination is effective. When transmitted light is used, filter to cut off long wavelength side is required for eliminating background.
 Ex  Effcy filter evaluation
 SC-56  SC-60
 470nm      
 505nm      *  
 530nm      
■ DsRed-Express2R

★ Place pointer on above figure to see LED & Filter wavelength
Exhibits strong and stable expression in bacterial cells with minimal toxicity.

Clontech Laboratories, Inc.

Em/Ex peaks are close to each other but both lines are spread over wide wavelength and therefore easier to separate. Blue, Cyan, Green LED can be used for excitation and red filter should be effective for separation.
 Ex  Effcy filter evaluation
 SC-56  SC-60
 470nm      
 505nm      *  
 530nm      
■ Ethidium Bromide

★ Place pointer on above figure to see LED & Filter wavelength

  Dramatic Effect of Dichroic Filter Effect ⇒

The most widely used fluorescent reagent for band staining. It is treated as hazardous waste by many organizations and require special care for handling.

Wikipedia

It has Ex peak at both 300/518nm.
LED 505/530nm can be used for excitation but efficiency is not so high and strong light source and effective filter for eliminating background is required.
 Ex  Effcy filter evaluation
 SC-56  SC-60
 470nm      
 505nm      *  
 530nm      

LED illuminator is equipped with high quality compound dichroic filter

LED505-TR60W
■ FITC

★ Place pointer on above figure to see LED & Filter wavelength
A kind of fluorochrome widely used for microscopic observation. Often utilized for bloodstain search, dye tracking in the area of forensic medicine and serology

Kirkegaard & Perry Lab

Has Excitation peak at 490nm and can be excited with 505nm LED. However Ex/Em is close and difficult to separate absorption and excitation spectra. High quality filter may required for eliminating background.
 Ex  Effcy filter evaluation
 SC-56  SC-60
 470nm      
 505nm      *  
 530nm      
■ Gel GreenR

★ Place pointer on above figure to see LED & Filter wavelength
Green fluorescent nucleic acid dye designed to replace the highly toxic EtBr. Compare to EtBr, it is ultra-sensitive and very stable, both hydrologically and thermally. Very simple procedures for pre or post staining

cBiotium

Wavelength characteristic is similar to FITC, and so is filterworks.
 Ex  Effcy filter evaluation
 SC-56  SC-60
 470nm      
 505nm      *  
 530nm      
■ Gel RedR

★ Place pointer on above figure to see LED & Filter wavelength
Fluorescent nucleic acid dye designed to replace the highly toxic EtBr. Compare to EtBr, it is ultra-sensitive and very stable, both hydrologically and thermally. Very simple procedures for pre or post staining.

cBiotium

Wavelength characteristic is similar to EtBr. Compare to EtBr, it has more abosrption around 520nm and thus
LED light is effective.
 Ex  Effcy filter evaluation
 SC-56  SC-60
 470nm      
 505nm      *  
 530nm      
■ GFP / EGFP

★ Place pointer on above figure to see LED & Filter wavelength
GFP enables real time detection without damaging cell. GFP also demonstrate its function as fusion protein as well (GFP tag) Widely used as a tool to locate protein which is taking part in signal transduction inside the cell.

GE healthcare

Em/Ex peaks are close to each other but Em is spread over to short wavelength side and therefore Blue LED should be effective for excitation
 Ex  Effcy filter evaluation
 SC-56  SC-60
 470nm      
 505nm      *  
 530nm      
■ mCherryR

★ Place pointer on above figure to see LED & Filter wavelength
Red fluorescent proteins which is derive from a protein isolated from Discosoma sp.Utilizes for fusion expression with target protein, promoter activity monitoring and bacteria expression.
Takes only short time for expression and also very stable.


Clontech Laboratories, Inc

Em/Ex peaks are close to each other but Em is spread over to short wavelength side and therefore Cyan and Green LED can be used for excitation. Excitation with Green LED and separation with red filter is effective.
 Ex  Effcy filter evaluation
 SC-56  SC-60
 470nm      
 505nm      *  
 530nm      
■ TagRFP

★ Place pointer on above figure to see LED & Filter wavelength
Generated from the wild-type RFP from sea anemone entacmaea quadricolor.
It is brightest monomeric red fluorescent protein available so far. Mammalian cells transiently transfected with TagRFP expression vectors produce bright fluorescence in 10-12 hrs after transfection.


Evrogen JSC

Cyan and especially Green LED has high excitation efficiency. Em is too close with Green LED, and background may appear with SC-56.Should be able to separate with SC-60 since Em is spread over to long wavelength side.
 Ex  Effcy filter evaluation
 SC-56  SC-60
 470nm      
 505nm      *  
 530nm      
■ SYBR GoldR

★ Place pointer on above figure to see LED & Filter wavelength

Life Technology

Relatively easy to excite with Blue and Cyan LED. Easy to separate since Ex/Em peak has some space between them.

 Ex  Effcy filter evaluation
 SC-56  SC-60
 470nm      
 505nm      *  
 530nm      
■ SYBR GreenR

★ Place pointer on above figure to see LED & Filter wavelength
Asymmetrical cyanine dye used as a nucleic acid stain in molecular biology.
It binds to double helix DNA and can be replacement for EtBr as it is safer to work
However any small molecule capable of binding DNA with high affinity is a possible carcinogen, including SYBR Green.


Life Technology

Ex is similar to SYBR Gold, however Em is adjacent to Ex and bit harder to separate
 Ex  Effcy filter evaluation
 SC-56  SC-60
 470nm      
 505nm      *  
 530nm      
■ SYPRO OrangeR

★ Place pointer on above figure to see LED & Filter wavelength
Provides fast, simple, sensitive staining of proteins. has very little protein-to-protein variation in staining. Enables long lasting an stable emission. With low background, it requires less time for cleaning gel. Detection with immunoreagent is also possible with western blotting.

GE healthcare

Ex wavelength is spread over to wide range and thus both Blue and Cyan LED are highly effective for excitation.
With enough space between Ex and Em,
it is relatively easy to separate as well.
 Ex  Effcy filter evaluation
 SC-56  SC-60
 470nm      
 505nm      *  
 530nm      
■ SYPRO RedR

★ Place pointer on above figure to see LED & Filter wavelength
For labeling the cellular tissue under the fluorescence microscope and for other studies of cell biology. It holds sulfonic acid which is directly connected to the fluorescent group. Especially bright with rhodamine derivative. The reagent with great light stability.

GE healthcare

Ex wavelength is spread over to wide range and thus both blue and cyan are effective for excitation. Most emission can transmit both SC-56 and SC-60 filter and so it should be relatively easy to separate as well.
 Ex  Effcy filter evaluation
 SC-56  SC-60
 470nm      
 505nm      *  
 530nm      
■ SYPRO RubyR

★ Place pointer on above figure to see LED & Filter wavelength
For labeling the cellular tissue under the fluorescence microscope and for other studies of cell biology. It holds sulfonic acid which is directly connected to the fluorescent group. Especially bright with rhodamine derivative. The reagent with great light stability.

GE healthcare

With wide space between the peaks of
Ex and Em, it is considered very easy reagent for separation. Blue LED is most effective for excitation and Cyan LED is also effective.
 Ex  Effcy filter evaluation
 SC-56  SC-60
 470nm      
 505nm      *  
 530nm      
* ⇒ Long cut filter may be required to eliminate background


 ** Please refer reagent list for Ex/Em info of other reagent ⇒  

Notice : The guide is reference purpose only. It does not gurantee any visibility of certain emission.
       Result may vary depends on such factors as LED brightness, filter quality, and camera spec.