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@@Filter Guide for Various Fluorescent Reagent (Ex/Em)


List of Reagents E Ex/Em wavelengths E LED light source E Filterwork
Following are list of most popular reagents we often receive inquiry.
Click Spectral button for filterwork guide of 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 mCitrine 516 529
Cy3 550 570 Midori Green 490 290 530
DsRed 557 579 SYBR Gold 495 540
EtBr 300 518 605 SYBR Green I 498 522
FITC 490 525 SYPRO Orange 470 570
Flamingo 512 271 535 SYPRO Red 550 300 630
Gel Green 250 500 530 SYPRO Ruby 280 450 620
Gel Red 270 510 600 TagRFP 555 583
GFP 488 507 tdTomato 554 581
mCherry 580 610 YFP 513 527
Ex/Em list for other reagents
Notice : The guide is reference purpose only. It does not guarantee visibility of
any certain emission. Result may vary depends on such various factors as
LED brightness, filter quality, and camera specifications, etc.
*  Long cut filter may required to eliminate background




Place pointer above and see LED/Filter wavelengths
For cellular labeling in fluorescence microscopy and in cell biology.
Retains sulfonic acid which is
directly connected to the fluorescent. Especially bright with rhodamine derivative. The reagent with great
light stability
Life Technology
Em/Ex are too close and difficult to separate absorption & excitation spectra. 505nm/SC-56 pair is effective. Filter to cut long
wavelength side is required to eliminate background when used
with transmitted light source.
Ex Effcy Filter evaluation
SC-56 SC-60
470nm
505nm *
530nm





Place pointer above and see LED/Filter wavelengths
In addition to such biological assays as immunocytochemistry, Cyanine
dye fluorescence reagent is often
used for labelling RNA.
Mirus Bio LLC
Em/Ex are close and difficult to separate absorption & excitation spectra. 505nm & SC-56 pair is effective. Filter to cut long
wavelength side is required to eliminate background when used
with transmitted light source.
Ex Effcy Filter evaluation
SC-56 SC-60
470nm
505nm *
530nm





Place pointer above and see LED/Filter wavelengths
Exhibits strong and stable
expression in bacterial cells
with minimal toxicity.
Clontech Laboratories, Inc
Em/Ex peaks are close but both lines are spread over wide wavelength and easier to separate. Blue, Cyan, Green LED can be used for excitation and
red filter is effective for separation.
Ex Effcy Filter evaluation
SC-56 SC-60
470nm
505nm *
530nm





Place pointer above and see LED/Filter wavelengths
The most widely used reagent for band staining. Treated as hazardous waste by many organizations and require special care for handling.
Wikipedia
It has Ex peak at both 300/518nm.
LED505/530nm can be used but efficiency is not high. High power light source and effective filter is required to eliminate background.
Ex Effcy Filter evaluation
SC-56 SC-60
470nm
505nm *
530nm



Dramatic Effect of
Dichroic Filter Effect

* Mecan LED transilluminator is equipped with high quality dichroic
filter
LC525

LED505-TR60W






Place pointer above and see LED/Filter wavelengths
A type 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
Ex peak at 490nm and can be excited with 505nm LED. However Ex/Em is close and may require LED device with background eliminating features. High power blue LED with SC52 filter can also be effective.
Ex Effcy Filter evaluation
SC-56 SC-60
470nm
505nm *
530nm
* 470nm + SC52 pair is also effective





Place pointer above and see LED/Filter wavelengths
Low cost protein detection with simple protocol. High sensitivity and quantitatively enables identification
of protein by mass spectrometry. Compatible also with UV excitation.
Bio-Rad
Ex 512nm and LED505nm can be effective. LED device with background eliminating feature or high power
blue LED & SC52 pair can also be effective since Ex/Em is close.
Ex Effcy Filter evaluation
SC-56 SC-60
470nm
505nm *
530nm
* 470nm + SC52 pair is also effective





Place pointer above and see LED/Filter wavelengths
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
* 470nm + SC52 pair is also effective





Place pointer above and see LED/Filter wavelengths
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





Place pointer above and see LED/Filter wavelengths
Enables real time detection without damaging cell. It also demonstrate its function as fusion protein (GFP tag) Widely used as a tool to locate
protein which is taking part in
signal transaction inside the cell.
GE healthcare
Em/Ex peaks are close but Em is spread over to short wavelength
side and Blue LED is effective
Ex Effcy Filter evaluation
SC-56 SC-60
470nm
505nm @*
530nm





Place pointer above and see LED/Filter wavelengths
Red fluorescent proteins isolated
from Discosoma sp. Utilizes for fusion expression with target protein, promoter activity monitoring and bacteria expression. Takes short time for expression and stable as well.
Clontech Laboratories, Inc
Em/Ex peaks are close but Em is spread over to short wavelength side and Cyan or Green LED can be used. Excitation with Green LED and separation with red filter is effective.
Ex Effcy Filter evaluation
SC-56 SC-60
470nm
505nm @*
530nm





Place pointer above and see LED/Filter wavelengths
mCitrine is an empty vector and gene insertion can be achieved by LIC cloning protocol. mCitrine is highly versatile with its compatibility with many other vector as well as usability as single expression vector.
? ? ?
Ex 510m and 505nm LED is efficient. However Ex/Em is close and may require LED device with background eliminating features. High power blue LED with SC52 filter is also effective.
Ex Effcy Filter evaluation
SC-56 SC-60
470nm
505nm @*
530nm
* 470nm + SC52 pair is also effective





Place pointer above and see LED/Filter wavelengths
Ideal Nucleic acid staining reagent for prestained method. Low background and high S/N ratio makes safe alternative to the ethidium bromide.
Fast Gene
Ralatively easy to excite with Blue LED (470m). Easy to separate since Ex/Em peak has some space between them.
Ex Effcy Filter evaluation
SC-54 SC-60
470nm
505nm
530nm
* SC52 is also effective





Place pointer above and see LED/Filter wavelengths
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





Place pointer above and see LED/Filter wavelengths
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. LED 505 with background eliminating features or high power blue LED & SC52 pair is also effective.
Ex Effcy Filter evaluation
SC-56 SC-60
470nm
505nm @*
530nm
* 470nm + SC52 pair is also effective





Place pointer above and see LED/Filter wavelengths
Provides fast, simple, sensitive staining of proteins. Has very little protein-to-protein variation in staining. Enables long lasting
and stable emission. With low background, it requires less time
for cleaning gel. Detection with immunoreagent is also possible
with western blotting.
GE healthcare
Ex is spread over to wide range
and both Blue and Cyan LED are
highly effective for excitation.
With enough space between Ex&Em,
it is relatively easy to separate too
Ex Effcy Filter evaluation
SC-56 SC-60
470nm
505nm @*
530nm





Place pointer above and see LED/Filter wavelengths
For cellular labeling in fluorescence microscopy and in cell biology.
Retains sulfonic acid which is
directly connected to the fluorescent. Especially bright with rhodamine derivative. The reagent with great
light stability

GE healthcare
Ex is spread over to wide range and both blue and cyan are effective for excitation. Most emission can transmit both SC-56 & SC-60 and it
is relatively easy to separate as well.
Ex Effcy Filter evaluation
SC-56 SC-60
470nm
505nm @*
530nm





Place pointer above and see LED/Filter wavelengths
For cellular labeling in fluorescence microscopy and in cell biology.
Retains sulfonic acid which is
directly connected to the fluorescent. Especially bright with rhodamine derivative. The reagent with great
light stability
GE healthcare
With wide space between the peaks
of Ex & Em, it is considered easy reagent for separation. Blue LED
is most effective for excitation.
Cyan LED is also effective.
Ex Effcy Filter evaluation
SC-56 SC-60
470nm
505nm @*
530nm





Place pointer above and see LED/Filter wavelengths
Generated from the wild-type RFP from sea anemone entacmaea quadricolor. It is brightest monomeric red fluorescent protein available. Mammalian cells transiently transfected with TagRFP expression vectors produce bright fluorescence
in 10-12 hrs after transfection.
Evrogen JSC
Cyan, or Green LED is effective for excitation. Em is too close with Green, and background may appear with SC-56 but sould 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





Place pointer above and see LED/Filter wavelengths
Exceptionally bright red fluorescent protein (2.5x brighter than EGFP) It is easily detected as deep as 1 cm below the surface, and ideal for live animal imaging studies. High cohesion stability make it suitable to produce protein N-, C- fusion protein too.
Takara Bio
Wavelength characteristics is similar to Tag RFP and Em/Ex are close.
Cyan LED & shortpass filters pair or Green LED & SC-60 filter pair should be used to eliminate background.
Ex Effcy Filter evaluation
SC-56 SC-60
470nm
505nm @*
530nm





Place pointer above and see LED/Filter wavelengths
YFP is synthetic non-aequorea fluorescent proteins that can be amplified/cut by various PCR and cloned into any other expression vector of choice. These vectors can also be used as expression vectors or as positive controls and allow monitoring of inducible protein expression.
Cosmo Bio
Ex peak at 512nm and can be excited with 505nm LED. However Ex/Em is close and may require LED device with background eliminating features. High power blue LED with SC52 filter can also be effective.
Ex Effcy Filter evaluation
SC-56 SC-60
470nm
505nm @*
530nm
* 470nm + SC52 pair is also effective



*  Long cut filter may required to eliminate background
Ex/Em list for other reagents
Notice : The guide is reference purpose only. It does not guarantee visibility of
any certain emission. Result may vary depends on such various factors as
LED brightness, filter quality, and camera specifications, etc.