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AAT Bioquest

Rhodamines and Rhodamine Derivatives

Rhodamines are xanthene derivatives structurally related to fluorescein, but with additional chemical substitutions that shift their excitation and emission spectra to longer wavelengths. The most widely used rhodamines include rhodamine 110, the parent rhodamine, tetramethylrhodamine (such as TAMRA or TRITC) and X-rhodamines (such as Texas Red®). In comparison to coumarins and cyanines, the fluorescence of rhodamines are generally more resistant to photodegradation and insensitive to pH changes. However, rhodamine conjugates are particularly susceptible to self-quenching. When more than two or three fluorophores are covalently attached to a target molecule, such as an antibody or nucleic acid, there is considerable loss in fluorescence.

 

 

Spectral Properties


Tetramethylrhodamine spectra
Excitation and emission spectra of tetramethylrhodamine (TRITC).

Table 1. Spectral properties of tetramethylrhodamine (TRITC).

Specification
Value
Laser Line:488/532 nm laser
Filter Set:TRITC
Ex (nm):552 nm
Em (nm):578 nm
Extinction Coefficient (ε):85,000 cm-1M-1
Quantum Yield (Φ):0.1




iFluor® 555 - a superior alternative to Cy3®, TRITC and Alexa Fluor® 555


Jurakt cells
Jurakt cells stained with Annexin V-iFluor® 555 conjugate.
The iFluor® 555 dye, which is spectrally similar to Cy3®, TRITC and Alexa Fluor® 555, produces significantly brighter and more photostable conjugates with pH-insensitive fluorescence over a wide molar range (pH 3 to 11). Ideal for imaging low-abundance targets with greater sensitivity, iFluor® 555 dye molecules can be conjugated to biomolecules, in particular antibodies and proteins, at high molar ratios with minimal self-quenching. These conjugates are well-suited for imaging and are particularly useful in multicolor applications, such as flow cytometry and fluorescence microscopy, without compromising the performance of blue and far-red fluorescent probes.

iFluor 555™ dyes are readily available in a variety of reactive dye forms and protein labeling kits allowing you to easily produce antibody conjugates or probes 'in-house'. Also available are iFluor 555™ dyes conjugated to antibodies, annexin V, phalloidin, amplification substrates (e.g. labeled-tyramides, labeled-styramides) and other biomolecules for cellular imaging and detection assays requiring increased sensitivity.

Learn More

 

Table 2. Available iFluor® 555 Reactive Dyes for Labeling Biomolecules.

Product
Chemical Reactivity
Reactive Moiety
Ex (nm)
Em (nm)
ε1
Unit Size
Cat No.
iFluor® 555 succinimidyl esterAmineSuccinimidyl Ester5575701000001 mg1028
iFluor® 555 maleimideThiolMaleimide5575701000001 mg1063
iFluor® 555 alkyneAzideAlkyne5575701000001 mg1092
iFluor® 555 amineAldehyde, Carboxylic Acid, KetoneAmine5575701000001 mg1073
iFluor® 555 azideAlkyneAzide5575701000001 mg1093
iFluor® 555 hydrazideAldehyde, Carboxylic Acid, KetoneHydrazide5575701000001 mg1083
  1. ε = molar extinction coefficient at their maximum absorption wavelength (Units = cm-1M-1).

 

Rhodamine Products


We offer a broad selection of rhodamine products including reactive rhodamine and rhodamine derivatives for labeling antibodies, nucleic acids and other biomolecules, conjugates, indicators, and substrates for detecting enzymatic activity in cells, homogenates and solution.

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Table 4. Reference Standards For Bioconjugation


Table 5. Rhodamine Precursors


 

Rhodamine Reactive Dyes


Reactive fluorescent rhodamine and rhodamine derivatives are widely used to covalently label biomolecules such as antibodies, carbohydrates, nucleic acids, proteins, peptides and oligonucleotides. Resulting in intensely bright and photostable orange fluorescence, rhodamine conjugates are often used as substrates for detecting enzymatic activity or to provide contrast in multicolored fluorescence investigation when combined with blue, green or far-red fluorescent probes. Rhodamine and rhodamine derivatives are readily available in a variety of dye-labeling chemistries, these include:
  • Succinimidyl esters/NHS esters - for labeling free amines (-NH2) on antibodies, proteins, peptides, amine-modified oligonucleotides and other biomolecules
  • Maleimides - for labeling sulfhydryl groups on antibodies, proteins and peptides
  • Azides - for labeling ethylene groups via click chemistry methods
  • Alkynes - for labeling azides via click chemistry methods
  • Hydrazides - for labeling aldehyde and ketone groups
  • Carboxylic acids - for labeling amines after pre-activation with carbodiimides or for Steglich esterification of alcohols
  • Amines - for labeling with various electrophilic compounds such as activated esters
  • Tetrazines - for labeling trans-cyclooctenes (TCO) compounds via Inverse-Electron-Demands Diels-Alder reaction
  • Phosphoramidite - for labeling oligonucleotides
 

Table 7. Available Reactive Rhodamine and Rhodamine Derivatives for Labeling Biomolecules.

Product
Chemical Reactivity
Reactive Moiety
Ex (nm)
Em (nm)
ε1
Unit Size
Cat No.
5-CR110 *Single isomer*Amines after pre-activation with carbodiimidesCarboxylic Acid499525840005 mg322
6-CR110 *Single isomer*Amines after pre-activation with carbodiimidesCarboxylic Acid499525840005 mg323
5(6)-CR6G [5-(and 6)-Carboxyrhodamine 6G] 5225469400025 mg330
5(6)-CR6G, SE *Mixed isomers*AmineSuccinimidyl Ester5225469400010 mg340
5(6)-CR6G, carboxylic acidAmines after pre-activation with carbodiimidesCarboxylic Acid5225469400010 mg340
5(6)-Carboxyrhodamine 6G cadaverineLabels proteins by transaminidationCadaverine522546940001 mg343
5(6)-Carboxyrhodamine 6G ethylenediamineAldehyde, Carboxylic Acid, KetoneEthylenediamine522546940001 mg344
5-CR6G, maleimideThiolMaleimide5255481160001 mg353
5-CR6G, SE *Single isomers*AmineSuccinimidyl Ester522546940005 mg345
5-CR6G, carboxylic acid *Single isomers*Amines after pre-activation with carbodiimidesCarboxylic Acid5225469400010 mg331
6-CR6G, SE *Single isomers*AmineSuccinimidyl Ester522546940001 mg342
6-CR6G, carboxylic acid *Single isomers*Amines after pre-activation with carbodiimidesCarboxylic Acid5225469400010 mg332
5(6)-TAMRA, SE *CAS 246256-50-8*AmineSuccinimidyl Ester5525789000025 mg370
5(6)-TAMRA, Maleimide *Mixed isomers*ThiolMaleimide552578900005 mg412
5(6)-TAMRA, carboxylic acid *CAS 98181-63-6*Amines after pre-activation with carbodiimidesCarboxylic Acid55257890000100 mg360
5(6)-TAMRA, ethylenediamineAldehyde, Carboxylic Acid, KetoneEthylenediamine5525789000025 mg354
5-TAMRA, SE *CAS 150810-68-7*AmineSuccinimidyl Ester552578900005 mg373
5-TAMRA, azideAlkyneAzide552578900005 mg486
5-TAMRA, alkyneAzideAlkyne552578900005 mg487
5-TAMRA, Maleimide *CAS 154480-30-5*ThiolMaleimide552578900001 mg421
5-TAMRA, carboxylic acid *CAS 91809-66-4*Amines after pre-activation with carbodiimidesCarboxylic Acid5525789000010 mg363
5-TAMRA, ethylenediamineAldehyde, Carboxylic Acid, KetoneEthylenediamine552578900005 mg358
6-TAMRA, SE *CAS 150810-69-8**AmineSuccinimidyl Ester552578900005 mg376
6-TAMRA, azideAlkyneAzide552578900005 mg490
6-TAMRA, alkyneAzideAlkyne552578900005 mg491
6-TAMRA, Maleimide *CAS 174568-68-4*ThiolMaleimide552578900001 mg419
6-TAMRA, carboxylic acid *CAS 91809-67-5*Amines after pre-activation with carbodiimidesCarboxylic Acid5525789000010 mg366
6-TAMRA, ethylenediamineAldehyde, Carboxylic Acid, KetoneEthylenediamine552578900005 mg359
6-TAMRA CPG *1000 Å*Oligonucleotide 3' end labeling 552578900005 mg358
5-TRITC [Tetramethylrhodamine-5-isothiocyanate]Amine 5445701000001 mg415
6-TRITC [Tetramethylrhodamine-6-isothiocyanate]Amine 5445701000001 mg417
5(6)-ROX, SE *Mixed isomers*AmineSuccinimidyl Ester5786048200025 mg390
5(6)-ROX, carboxylic acid *CAS 198978-94-8*Amines after pre-activation with carbodiimidesCarboxylic Acid57860482000100 mg380
5-ROX, SE *CAS 209734-74-7*AmineSuccinimidyl Ester5786048200025 mg389
5-ROX, carboxylic acid *CAS 216699-35-3*Amines after pre-activation with carbodiimidesCarboxylic Acid57860482000100 mg381
6-ROX, SE *CAS 216699-36-4*AmineSuccinimidyl Ester578604820005 mg392
6-ROX, azideAlkyneAzide578604820005 mg494
6-ROX, alkyneAzideAlkyne578604820005 mg495
6-ROX C2 MaleimideThiolMaleimide578604820001 mg419
6-ROX, carboxylic acid *CAS 216699-35-3*Amines after pre-activation with carbodiimidesCarboxylic Acid5786048200025 mg382
6-ROXtra™ acid 578595 25 mg386
6-ROXtra™ SEAmineSuccinimidyl Ester578595 5 mg397
Bodi Fluor™ R6G NHS Ester [equivalent to Bodipy® R6G, NHS Ester]AmineSuccinimidyl Ester5285471160005 mg703
LRB Red™ SEAmineSuccinimidyl Ester558575 10 mg471
Lissamine Rhodamine B Sulfonyl Chloride *CAS 62796-29-6*AmineSulfonyl Chloride558575 100 mg470
Texas Red-X, succinimidyl ester *Mixed isomers* *CAS 216972-99-5*AmineSuccinimidyl Ester5866031160005 mg475
Texas Red-X, succinimidyl ester *Single isomer* *CAS 199745-67-0*AmineSuccinimidyl Ester5866031160005 mg474
Sulforhodamine 101 sulfonyl chloride [Texas Red®]AmineSulfonyl Chloride58660311600010 mg480
Texas Red® hydrazideAldehyde, Carboxylic Acid, KetoneHydrazide5866031160005 mg481
Texas Red® maleimideThiolMaleimide5866031160005 mg483
Texas Red® azideAlkyneAzide5866031160005 mg484
Texas Red® cadaverineLabels proteins by transaminidationCadaverine5866031160005 mg482
Texas Red® alkyneAzideAlkyne5866031160005 mg485
  1. ε = molar extinction coefficient at their maximum absorption wavelength (Units = cm-1M-1).

 

Rhodamine-Labeled Enzyme Substrates


Rhodamine and rhodamine derivatives are frequently used to prepare fluorogenic substrates for detecting enzymatic activity in cells and solutions. When hydrolyzed by the enzyme of interest, rhodamine-modified substrates exhibit an increase in fluorescence intensity that is linearly proportional to enzyme activity. AAT Bioquest provides a large assortment of rhodamine modified substrates for detecting activity from caspases, proteases, peptidases, phosphatases and more.
 

Table 8. Available Fluorescein-Labeled Fluorogenic Substrates.

Product
Biological Function
Ex (nm)
Em (nm)
Unit Size
Cat No.
FDG [Fluorescein di-beta-D-galactopyranoside]β-galactosidase substrate4985171 g14001
FDP [Fluorescein diphosphate, tetraammonium salt] *CAS 217305-49-2*Phosphatase Substrate4985175 mg11600
FCB [Fluorescein di-beta-D-cellobioside]Cellulase (e.g. β-glucosidases, endoglucanases and exoglucanases) Substrate4985171 mg14025
Casein, FITC-conjugatedGeneric Protease Substrate4985175 mg13440
FAM-VAD-FMKPolycaspase Inhibitor (Caspases 1, 2, 3, 6, 8, 9, or 10)49351725 Tests13470
FITC-C6-DEVD-FMKCaspase 3 Substrate491516100 µg13408
FITC-C6-D(OMe)E(OMe)VD(OMe)-FMKCaspase 3 Substrate491516100 µg13417
FITC-C6-LEHD-FMKCaspase 9 Substrate491516100 µg13409
FDGlcU [Fluorescein di-beta-D-glucuronide]β-Glucuronidase Substrate4985171 mg14002
Glucose-UDP-Fluorescein ConjugateGlucosyltransferase Substrate498517100 µg11705
Glucose-UDP-(PEG)6-Fluorescein ConjugateGlucosyltransferase Substrate498517100 µg11706

 

Rhodamine-Labeled Conjugates


Rhodamine-labeled conjugates have a wide range of applications in bio-imaging and bio-labelling due to their bright orange fluorescence and excellent photostability. Rhodamine derivatives can be labeled to antibodies, streptavidin, phalloidin, annexin v, dextran and other biomolecules to study apoptosis, label organelles or improve assay sensitivity by amplifying weak signals.
 

Table 9. Available Rhodamine-Labeled Fluorogenic Conjugates.

Product
Biological Function
Ex (nm)
Em (nm)
Unit Size
Cat No.
TRITC Goat Anti-human IgG (H+L) AntibodyIndirectly detects human primary antibodies5445701 mg50209
TRITC Goat Anti-human IgG (H+L) Antibody *Cross Adsorbed*Indirectly detects human primary antibodies5445701 mg50211
Phalloidin-Tetramethylrhodamine ConjugateLabels actin filaments (F-actin)552578300 Tests23102
Annexin V, TRITC LabeledDetects phosphatidylserine on the cell surface544570100 Tests20031
TRITC–Dextran Conjugate (average MW = ~20K)Cardiovascular, Microcirculation, Perfusion, Cell Membrane Flux Tracer54457025 mg21706
TRITC–Dextran Conjugate (average MW = ~70K)Cardiovascular, Microcirculation, Perfusion, Cell Membrane Flux Tracer54457025 mg21708
TRITC–Dextran Conjugate (average MW = ~150K)Cardiovascular, Microcirculation, Perfusion, Cell Membrane Flux Tracer54457025 mg21710
Texas Red®-streptavidin conjugateCardiovascular, Microcirculation, Perfusion, Cell Membrane Flux Tracer5866031 mg16911
Texas Red® goat anti-mouse IgG (H+L)Indirectly detects mouse primary antibodies5866031 mg16853
Texas Red® goat anti-mouse IgG (H+L) *Cross Adsorbed*Indirectly detects mouse primary antibodies5866031 mg16861
Texas Red® goat anti-rabbit IgG (H+L)Indirectly detects rabbit primary antibodies5866031 mg16869
Texas Red® goat anti-rabbit IgG (H+L) *Cross Adsorbed*Indirectly detects rabbit primary antibodies5866031 mg16877
PE/Texas Red® goat anti-human IgG (H+L)Indirectly detects human primary antibodies5866031 mg50249
PE/Texas Red® goat anti-human IgG (H+L) *Cross Adsorbed*Indirectly detects human primary antibodies5866031 mg50251

 

Rhodamine-Derived Indicators


Rhodamine-derived indicators are important tools in biochemical and cellular studies. They have been used in all major types of fluorescence imaging applications including flow cytometry, fluorescence microscopy and fluorescence plate-based assays. AAT Bioquest offers a wide range of rhodamine-derived indicators for measuring cellular pH, calcium concentrations, cell proliferation, cell viability and more.
 

Table 10. Available Rhodamine-Derived Indicators.

Product
Biological Function
Ex (nm)
Em (nm)
Unit Size
Cat No.
Rhodamine B *CAS 81-88-9* 546568100 mg69
Rhodamine 6G *CAS 989-38-8* 525548100 mg70
Sulforhodamine B *CAS 3520-42-1* 5595771 g72
Sulforhodamine G *CAS 5873-16-5* 5315521 g73
Rhodamine 123 *CAS 62669-70-9*Live Cell Mitochondria Indicator50852825 mg22210
Dihydrorhodamine 123 *CAS 109244-58-8*Redox Indicator50852810 mg15206
Rhodamine B, hexyl ester, perchlorateLive Cell Mitochondria Indicator54656810 mg22211
TMRM [Tetramethylrhodamine methyl ester] *CAS#: 115532-50-8*Mitochondrial Membrane Potential Indicator55257425 mg22221
TMRE [Tetramethylrhodamine ethyl ester] *CAS#: 115532-52-0*Mitochondrial Membrane Potential Indicator55257425 mg22220
TRITC-DHPEPhospholipid Tracer5445701 mg23301
Texas Red-DHPE *CAS 187099-99-6*Phospholipid Tracer5866031 mg23300
Rhod-2, AM *UltraPure Grade* *CAS 145037-81-6*Cell-Permeable Calcium Indicator (Kd = 570 nM)5535771 mg21062
Rhod-2, trisodium saltCell-Impermeable Calcium Indicator (Kd = 570 nM)5535771 mg21068
Rhod-2, tripotassium saltCell-Impermeable Calcium Indicator (Kd = 570 nM)5535771 mg21067
Rhod-2 dextran conjugate *Low affinity with MW 10,000*Cell-Impermeable Calcium Indicator (Kd = 3.8 µM)5535771 mg20451
Rhod-4™, AMCell-Permeable Calcium Indicator (Kd = 451 nM)5235511 mg21120
Rhod-4™, sodium saltCell-Impermeable Calcium Indicator (Kd = 451 nM)5235511 mg21118
Rhod-4™, potassium saltCell-Impermeable Calcium Indicator (Kd = 451 nM)5235511 mg21119
Rhod-5N, AMCell-Permeable Calcium Indicator (Kd = 320 µM)5575801 mg21070
Rhod-5N, tripotassium saltCell-Impermeable Calcium Indicator (Kd = 320 µM)5575801 mg21072
Rhod-FF, AMCell-Permeable Calcium Indicator (Kd = 19 µM)5535771 mg21077
Rhod-FF, tripotassium saltCell-Impermeable Calcium Indicator (Kd = 19 µM)5535771 mg21075