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iFluor® 594 Tyramide

Formalin-fixed, paraffin-embedded (FFPE) human lung tissue was labeled with anti-EpCAM mouse mAb followed by HRP-labeled goat anti-mouse IgG (Cat No. 16728). The fluorescence signal was developed using iFluor® 594 tyramide (Cat No. 45107) and detected with a TRITC/Cy3 filter set. Nuclei (blue) were counterstained with DAPI (Cat No. 17507).
Formalin-fixed, paraffin-embedded (FFPE) human lung tissue was labeled with anti-EpCAM mouse mAb followed by HRP-labeled goat anti-mouse IgG (Cat No. 16728). The fluorescence signal was developed using iFluor® 594 tyramide (Cat No. 45107) and detected with a TRITC/Cy3 filter set. Nuclei (blue) were counterstained with DAPI (Cat No. 17507).
<strong>Superior sensitivity with iFluor® 594 tyramide.</strong> HeLa cells were incubated with primary anti-tubulin antibodies followed by detection with HRP-Goat anti-Mouse IgG and iFluor® 594 tyramide (Left) or Alexa Fluor® 594 tyramide (Right). Fluorescence images were taken on a Keyence BZ-X710 fluorescence microscope equipped with a TRITC filter set.
Chemical structure for iFluor® 594 Tyramide.
Ordering information
Price ()
Catalog Number45107
Unit Size
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Additional ordering information
Telephone1-408-733-1055
Fax1-408-733-1304
Emailsales@aatbio.com
InternationalSee distributors
ShippingStandard overnight for United States, inquire for international
Physical properties
Molecular weight841.95
SolventDMSO
Spectral properties
Absorbance (nm)587
Correction Factor (260 nm)0.05
Correction Factor (280 nm)0.04
Extinction coefficient (cm -1 M -1)1800001
Excitation (nm)588
Emission (nm)604
Quantum yield0.531
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure
UNSPSC12171501
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iFluor® 594 succinimidyl ester
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iFluor® Ultra 594 succinimidyl ester
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OverviewpdfSDSpdfProtocol


Molecular weight
841.95
Absorbance (nm)
587
Correction Factor (260 nm)
0.05
Correction Factor (280 nm)
0.04
Extinction coefficient (cm -1 M -1)
1800001
Excitation (nm)
588
Emission (nm)
604
Quantum yield
0.531
For many immunohistochemical (IHC) applications, the traditional enzymatic amplification procedures are sufficient for achieving adequate antigen detection. However, several factors limit the sensitivity and utility of these procedures. Tyramide signal amplification (TSA) has proven to be a particularly versatile and powerful enzyme amplification technique with improved assay sensitivity. TSA is based on the ability of HRP, in the presence of low concentrations of hydrogen peroxide, to convert labeled tyramine-containing substrate into an oxidized, highly reactive free radical that can covalently bind to tyrosine residues at or near the HRP. To achieve maximal IHC detection, tyramine is prelabeled with a fluorophore. The signal amplification conferred by the turnover of multiple tyramide substrates per peroxidase label translates ultrasensitive detection of low-abundance targets and the use of smaller amounts of antibodies and hybridization probes. In immunohistochemical applications, sensitivity enhancements derived from TSA method allow primary antibody dilutions to be increased to reduce nonspecific background signals, and can overcome weak immunolabeling caused by suboptimal fixation procedures or low levels of target expression. iFluor 594 tyramide contains the bright iFluor 594 that can be readily detected with the standard Texas Red filter set. It is an excellent replacement for Alexa Fluor® 594 tyramide (Alexa Fluor® is the trade mark of ThermoFisher) or other spectrally similar fluorescent tyramide conjugates or TSA reagents (such as fluorescein tyramide).

Calculators


Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of iFluor® 594 Tyramide to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

0.1 mg0.5 mg1 mg5 mg10 mg
1 mM118.772 µL593.859 µL1.188 mL5.939 mL11.877 mL
5 mM23.754 µL118.772 µL237.544 µL1.188 mL2.375 mL
10 mM11.877 µL59.386 µL118.772 µL593.859 µL1.188 mL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

Mass (Calculate)Molecular weightVolume (Calculate)Concentration (Calculate)Moles
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Spectrum


Open in Advanced Spectrum Viewer
spectrum

Spectral properties

Absorbance (nm)587
Correction Factor (260 nm)0.05
Correction Factor (280 nm)0.04
Extinction coefficient (cm -1 M -1)1800001
Excitation (nm)588
Emission (nm)604
Quantum yield0.531

Product family


NameExcitation (nm)Emission (nm)Extinction coefficient (cm -1 M -1)Quantum yieldCorrection Factor (260 nm)Correction Factor (280 nm)
iFluor® 488 tyramide4915167500010.910.210.11
iFluor® 594 maleimide58860418000010.5310.050.04
iFluor® 594 Styramide *Superior Replacement for Alexa Fluor 594 tyramide*58860418000010.5310.050.04
iFluor® 555 Tyramide55757010000010.6410.230.14
iFluor® 647 Tyramide65667025000010.2510.030.03
iFluor® 350 Tyramide3454502000010.9510.830.23
iFluor® 546 Tyramide54155710000010.6710.250.15
iFluor® 568 Tyramide56858710000010.5710.340.15
iFluor® 594 TCO58860418000010.5310.050.04
iFluor® 594 Tetrazine58860418000010.5310.050.04
iFluor® 633 tyramide64065425000010.2910.0620.044
iFluor® 430 Tyramide *Superior Replacement for Opal 480*4334984000010.7810.680.3
iFluor® 450 Tyramide *Superior Replacement for Opal 480*4515024000010.8210.450.27
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References


View all 14 references: Citation Explorer
Time Gated Luminescence Imaging of Immunolabeled Human Tissues.
Authors: Chen, Ting and Hong, Rui and Magda, Darren and Bieniarz, Christopher and Morrison, Larry and Miller, Lawrence W
Journal: Analytical chemistry (2017): 12713-12719
Whole Mount RNA-FISH on Ovules and Developing Seeds.
Authors: Bleckmann, Andrea and Dresselhaus, Thomas
Journal: Methods in molecular biology (Clifton, N.J.) (2017): 159-171
Reliable protocols for whole-mount fluorescent in situ hybridization (FISH) in the pea aphid Acyrthosiphon pisum: a comprehensive survey and analysis.
Authors: Chung, Chen-yo and Cook, Charles E and Lin, Gee-way and Huang, Ting-Yu and Chang, Chun-che
Journal: Insect science (2014): 265-77
Quantum dot-based FRET for sensitive determination of hydrogen peroxide and glucose using tyramide reaction.
Authors: Huang, Xiangyi and Wang, Jinjie and Liu, Heng and Lan, Tao and Ren, Jicun
Journal: Talanta (2013): 79-84
NIR-labeled perfluoropolyether nanoemulsions for drug delivery and imaging.
Authors: O'Hanlon, Claire E and Amede, Konjit G and O'Hear, Meredith R and Janjic, Jelena M
Journal: Journal of fluorine chemistry (2012): 27-33
Anisotropic magnetic porous assemblies of oxide nanoparticles interconnected via silica bridges for catalytic application.
Authors: Wacker, Josias B and Parashar, Virendra K and Gijs, Martin A M
Journal: Langmuir : the ACS journal of surfaces and colloids (2011): 4380-5
Methoxychlor and estradiol induce oxidative stress DNA damage in the mouse ovarian surface epithelium.
Authors: Symonds, Daniel A and Merchenthaler, Istvan and Flaws, Jodi A
Journal: Toxicological sciences : an official journal of the Society of Toxicology (2008): 182-7
mRNA-targeted fluorescent in situ hybridization (FISH) of Gram-negative bacteria without template amplification or tyramide signal amplification.
Authors: Coleman, James R and Culley, David E and Chrisler, William B and Brockman, Fred J
Journal: Journal of microbiological methods (2007): 246-55
Multiplex fluorescent in situ hybridization in zebrafish embryos using tyramide signal amplification.
Authors: Clay, Hilary and Ramakrishnan, Lalita
Journal: Zebrafish (2005): 105-11
Oligo(dA-dT)-dependent signal amplification for the detection of proteins in cells.
Authors: Hanaki, Ken-ichi and Ohka, Seii and Yamamoto, Kenji and Nomoto, Akio and Yoshikura, Hiroshi
Journal: BioTechniques (2004): 856-60, 862-3