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iFluor® 568 Tyramide

Microtubules of fixed HeLa cells were labeled with anti-α tubulin mouse mAb followed by HRP-labeled goat anti-mouse IgG (Cat No. 16728). The fluorescence signal was developed using Alexa Fluor® 568 tyramide or iFluor® 568 tyramide (Cat No. 45106) and detected with a TRITC/Cy3 filter set. iFluor® 568 tyramide shows significantly higher fluorescence intensity than Alexa Fluor® 568 tyramide under the same conditions.
Microtubules of fixed HeLa cells were labeled with anti-α tubulin mouse mAb followed by HRP-labeled goat anti-mouse IgG (Cat No. 16728). The fluorescence signal was developed using Alexa Fluor® 568 tyramide or iFluor® 568 tyramide (Cat No. 45106) and detected with a TRITC/Cy3 filter set. iFluor® 568 tyramide shows significantly higher fluorescence intensity than Alexa Fluor® 568 tyramide under the same conditions.
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® 568 tyramide (Cat No.  45106) or Alexa Fluor® 568 tyramide and detected with a TRITC/Cy3 filter set. Nuclei (blue) were counterstained with DAPI (Cat No. 17507).
Chemical structure for iFluor® 568 Tyramide.
Ordering information
Price ()
Catalog Number45106
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 weight813.89
SolventDMSO
Spectral properties
Correction Factor (260 nm)0.34
Correction Factor (280 nm)0.15
Extinction coefficient (cm -1 M -1)1000001
Excitation (nm)568
Emission (nm)587
Quantum yield0.571
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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OverviewpdfSDSpdfProtocol


Molecular weight
813.89
Correction Factor (260 nm)
0.34
Correction Factor (280 nm)
0.15
Extinction coefficient (cm -1 M -1)
1000001
Excitation (nm)
568
Emission (nm)
587
Quantum yield
0.571
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 568 tyramide contains the bright iFluor 568 that can be readily detected with the standard ROX filter set. It is an excellent replacement for Alexa Fluor® 568 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® 568 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 mM122.867 µL614.334 µL1.229 mL6.143 mL12.287 mL
5 mM24.573 µL122.867 µL245.733 µL1.229 mL2.457 mL
10 mM12.287 µL61.433 µL122.867 µL614.334 µL1.229 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


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spectrum

Spectral properties

Correction Factor (260 nm)0.34
Correction Factor (280 nm)0.15
Extinction coefficient (cm -1 M -1)1000001
Excitation (nm)568
Emission (nm)587
Quantum yield0.571

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® 568 maleimide56858710000010.5710.340.15
iFluor® 568 Styramide *Superior Replacement for Alexa Fluor 568 tyramide*56858710000010.5710.340.15
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® 594 Tyramide58860418000010.5310.050.04
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 50 references: Citation Explorer
Microwaving and Fluorophore-Tyramide for Multiplex Immunostaining on Mouse Adrenals - Using Unconjugated Primary Antibodies from the Same Host Species.
Authors: Lyu, Qiongxia and Zheng, Huifei Sophia and Laprocina, Karly and Huang, Chen-Che Jeff
Journal: Journal of visualized experiments : JoVE (2020)
Detection of Cytokine Receptors Using Tyramide Signal Amplification for Immunofluorescence.
Authors: Wang, Herui and Pangilinan, Ryan L and Zhu, Yan
Journal: Methods in molecular biology (Clifton, N.J.) (2020): 89-97
Procedural Requirements and Recommendations for Multiplex Immunofluorescence Tyramide Signal Amplification Assays to Support Translational Oncology Studies.
Authors: Parra, Edwin Roger and Jiang, Mei and Solis, Luisa and Mino, Barbara and Laberiano, Caddie and Hernandez, Sharia and Gite, Swati and Verma, Anuj and Tetzlaff, Michael and Haymaker, Cara and Tamegnon, Auriole and Rodriguez-Canales, Jaime and Hoyd, Clifford and Bernachez, Chantale and Wistuba, Ignacio
Journal: Cancers (2020)
Sensitive Multiplexed Fluorescent In Situ Hybridization Using Enhanced Tyramide Signal Amplification and Its Combination with Immunofluorescent Protein Visualization in Zebrafish.
Authors: Lauter, Gilbert and Söll, Iris and Hauptmann, Giselbert
Journal: Methods in molecular biology (Clifton, N.J.) (2020): 397-409
Optimization of prostate cancer cell detection using multiplex tyramide signal amplification.
Authors: Roy, Sounak and Axelrod, Haley D and Valkenburg, Kenneth C and Amend, Sarah and Pienta, Kenneth J
Journal: Journal of cellular biochemistry (2019): 4804-4812
Highly Sensitive Detection of PCV2 Based on Tyramide Signals and GNPL Amplification.
Authors: Zhang, Shouping and Hu, Bin and Xia, Xiaojing and Xu, Yanzhao and Hang, Bolin and Jiang, Jinqing and Hu, Jianhe
Journal: Molecules (Basel, Switzerland) (2019)
DNAzyme Catalyzed Tyramide Depositing Reaction for In Situ Imaging of Protein Status on the Cell Surface.
Authors: Xu, Lulu and Liu, Shengchun and Yang, Tiantian and Shen, Yifan and Zhang, Yuhong and Huang, Lizhen and Zhang, Lutan and Ding, Shijia and Song, Fangzhou and Cheng, Wei
Journal: Theranostics (2019): 1993-2002
Cancer immunophenotyping by seven-colour multispectral imaging without tyramide signal amplification.
Authors: Ijsselsteijn, Marieke E and Brouwer, Thomas P and Abdulrahman, Ziena and Reidy, Eileen and Ramalheiro, Ana and Heeren, A Marijne and Vahrmeijer, Alexander and Jordanova, Ekaterina S and de Miranda, Noel Fcc
Journal: The journal of pathology. Clinical research (2019): 3-11
Comparative Tyramide-FISH mapping of the genes controlling flavor and bulb color in Allium species revealed an altered gene order.
Authors: Khrustaleva, Ludmila and Kudryavtseva, Natalia and Romanov, Dmitry and Ermolaev, Aleksey and Kirov, Ilya
Journal: Scientific reports (2019): 12007
An ultrasensitive electrochemical immunosensor for procalcitonin detection based on the gold nanoparticles-enhanced tyramide signal amplification strategy.
Authors: Liu, Pei and Li, Chao and Zhang, Ruixuan and Tang, Qing and Wei, Jia and Lu, Yan and Shen, Pingping
Journal: Biosensors & bioelectronics (2019): 543-550