iFluor®647-PEG12-dUTP 1 mM in Tris Buffer (pH 7.5)

Fluorescence In Situ Hybridization of Cy5 and iFluor® 647-dUTP labelled Telomere probes in metaphase HeLa cells.

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Physical properties

Molecular weight	2293.72
Solvent	Water

Spectral properties

Correction Factor (260 nm)	0.03
Correction Factor (280 nm)	0.03
Correction Factor (656 nm)	0.0793
Extinction coefficient (cm ^-1 M ^-1)	250000¹
Excitation (nm)	656
Emission (nm)	670
Quantum yield	0.25¹

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Freeze (< -15 °C); Minimize light exposure
UNSPSC	12171501

Overview SDS Protocol

Molecular weight

2293.72

Correction Factor (260 nm)

0.03

Correction Factor (280 nm)

0.03

Correction Factor (656 nm)

0.0793

Extinction coefficient (cm ^-1 M ^-1)

250000¹

Excitation (nm)

656

Emission (nm)

670

Quantum yield

0.25¹

The dye-modified deoxyuridine 5'-triphosphates are one of the most common methods to produce dye-labeled DNA via conventional enzymatic incorporation methods such as reverse transcription, nick translation, random primed labeling, or PCR. This enzymatic fluorescence labeling method is widely used for both FISH probes and microarray-based experiments. This iFluor® 647-PEG12-dUTP conjugate has the same spectral properties as ChromaTide™ Alexa Fluor™ 647-dUTP. Its PEG12 spacer makes it much more effectively incorporated into DNAs by polymerase. It is used as a far-red fluorescence color with a SpectrumFRed™ filter set. It provides a bright signal that has high photostability and is not affected by pH. ChromaTide™ and Alexa Fluor™ are the trademarks of ThermoFisher, and SpectrumFRed™ is the trademark of Vysis.

Calculators

Common stock solution preparation

Table 1. Volume of Water needed to reconstitute specific mass of iFluor®647-PEG12-dUTP *1 mM in Tris Buffer (pH 7.5)* to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

	0.1 mg	0.5 mg	1 mg	5 mg	10 mg
1 mM	43.597 µL	217.987 µL	435.973 µL	2.18 mL	4.36 mL
5 mM	8.719 µL	43.597 µL	87.195 µL	435.973 µL	871.946 µL
10 mM	4.36 µL	21.799 µL	43.597 µL	217.987 µL	435.973 µL

Molarity calculator

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

Mass (Calculate)		Molecular weight		Volume (Calculate)		Concentration (Calculate)		Moles
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Spectrum

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Spectral properties

Correction Factor (260 nm)	0.03
Correction Factor (280 nm)	0.03
Correction Factor (656 nm)	0.0793
Extinction coefficient (cm ^-1 M ^-1)	250000¹
Excitation (nm)	656
Emission (nm)	670
Quantum yield	0.25¹

Product Family

Name	Excitation (nm)	Emission (nm)	Extinction coefficient (cm ^-1 M ^-1)	Quantum yield	Correction Factor (260 nm)	Correction Factor (280 nm)
iFluor®488-PEG12-dUTP 1 mM in Tris Buffer (pH 7.5)	491	516	75000¹	0.9¹	0.21	0.11
iFluor®555-PEG12-dUTP 1 mM in Tris Buffer (pH 7.5)	557	570	100000¹	0.64¹	0.23	0.14

Images

Figure 1. Fluorescence In Situ Hybridization of Cy5 and iFluor® 647-dUTP labelled Telomere probes in metaphase HeLa cells.

Figure 2. Chemical structure for iFluor®647-PEG12-dUTP *1 mM in Tris Buffer (pH 7.5)*.

References

View all 38 references: Citation Explorer

Loss of PDPK1 abrogates resistance to gemcitabine in label-retaining pancreatic cancer cells.
Authors: Li, Dandan and Mullinax, John E and Aiken, Taylor and Xin, Hongwu and Wiegand, Gordon and Anderson, Andrew and Thorgeirsson, Snorri and Avital, Itzhak and Rudloff, Udo
Journal: BMC cancer (2018): 772

dUTPs conjugated with zwitterionic Cy3 or Cy5 fluorophore analogues are effective substrates for DNA amplification and labelling by Taq polymerase.
Authors: Zasedateleva, Olga A and Vasiliskov, Vadim A and Surzhikov, Sergey A and Kuznetsova, Viktoriya E and Shershov, Valeriy E and Guseinov, Timur O and Smirnov, Igor P and Yurasov, Roman A and Spitsyn, Maksim A and Chudinov, Alexander V
Journal: Nucleic acids research (2018): e73

Mass-spectrometry analysis of modifications at DNA termini induced by DNA polymerases.
Authors: Smirnov, Igor P and Kolganova, Natalia A and Vasiliskov, Vadim A and Chudinov, Alexander V and Timofeev, Edward N
Journal: Scientific reports (2017): 6674

Separation of replication and transcription domains in nucleoli.
Authors: Smirnov, E and Borkovec, J and Kováčik, L and Svidenská, S and Schröfel, A and Skalníková, M and Švindrych, Z and Křížek, P and Ovesný, M and Hagen, G M and Juda, P and Michalová, K and Cardoso, M C and Cmarko, D and Raška, I
Journal: Journal of structural biology (2014): 259-66

A prenatally ascertained, maternally inherited 14.8 Mb duplication of chromosomal bands Xq13.2-q21.31 associated with multiple congenital abnormalities in a male fetus.
Authors: Sismani, C and Donoghue, J and Alexandrou, A and Karkaletsi, M and Christopoulou, S and Konstantinidou, A E and Livanos, P and Patsalis, P C and Velissariou, V
Journal: Gene (2013): 138-42

[Establishment of a hydrogel chip for high-throughput detection of Y chromosome microdeletions].
Authors: Li, You-Zhi and Chen, Zhi-Yao and Wang, Hui and Huang, Huan and Song, Qin-Xin and Zhou, Guo-Hua
Journal: Zhonghua nan ke xue = National journal of andrology (2012): 109-14

Selection of reliable reference genes for gene expression study in nasopharyngeal carcinoma.
Authors: Guo, Yi and Chen, Jia-xin and Yang, Shu and Fu, Xu-ping and Zhang, Zheng and Chen, Ke-he and Huang, Yan and Li, Yao and Xie, Yi and Mao, Yu-min
Journal: Acta pharmacologica Sinica (2010): 1487-94

Time-resolved three-dimensional molecular tracking in live cells.
Authors: Wells, Nathan P and Lessard, Guillaume A and Goodwin, Peter M and Phipps, Mary E and Cutler, Patrick J and Lidke, Diane S and Wilson, Bridget S and Werner, James H
Journal: Nano letters (2010): 4732-7

[Expression of cell cycle molecules in human azoospermic testes].
Authors: Yang, Bo and Yuan, Jian-lin and Gao, Xiao-kang and Qin, Wei-jun and Liu, Fei and Shao, Chen and Liu, He-liang and Kang, Fu-xia
Journal: Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology (2009): 393-5

[Expression of COX10 in human non-obstructive azoospermia testes].
Authors: Yang, Bo and Yuan, Jian-Lin and Gao, Xiao-Kang and Wang, He and Shao, Chen and Liu, He-Liang and Chen, Bao-Qi and Qin, Rong-Liang and Shao, Guo-Xing and Kang, Fu-Xia
Journal: Zhonghua nan ke xue = National journal of andrology (2009): 599-603