ReadiUse™ Preactivated PE-iFluor® 660 Tandem

Our preactivated PE-iFluor® 660 Tandem was premodified with our Buccutite™ FOL (provided). Your antibody (or other proteins) is modified with our Buccutite™ MTA (provided as free sample) to give MTA-modified protein (such as antibody). The MTA-modified protein readily reacts with FOL-modified PE-iFluor® 660 Tandem (provided) to give the desired PE-iFluor® 660 Tandem-antibody conjugate in much higher yield than the SMCC chemistry. In addition our preactivated PE-iFluor® 660 Tandem reacts with MTA-modified biopolymers at much lower concentrations than the SMCC chemistry.

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Additional ordering information

Telephone	1-800-990-8053
Fax	1-800-609-2943
Email	sales@aatbio.com
Quotation	Request
International	See distributors
Shipping	Standard overnight for United States, inquire for international

Physical properties

Solvent

Water

Spectral properties

Absorbance (nm)	566
Extinction coefficient (cm ^-1 M ^-1)	1960000
Excitation (nm)	565
Emission (nm)	695

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Refrigerated (2-8 °C); Minimize light exposure
UNSPSC	12171501

Overview SDS Protocol

Absorbance (nm)

566

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

1960000

Excitation (nm)

565

Emission (nm)

695

R-Phycoerythrin (PE) is isolated from red algae. Its primary absorption peak is at 565 nm with secondary peaks at 496 and 545 nm. Our RPE-iFluor® 660 Tandem is an excellent color for multicolor analysis of multiple targets with flow cytometry. It has a unique emission located between the emission Cy5 and that of Cy5.5, providing a new color for spectral flow cytometry. ReadiUse™ Preactivated PE-iFluor® 660 Tandem is an activated PE protein, and can be easily conjugated to antibodies with much higher conjugation yield than the conventional PE.

Components

Example protocol

AT A GLANCE

Important PE-iFluor™ 660 Tandem was premodified with our Buccutite™ FOL. Your antibody (or other proteins) is modified with our Buccutite™ MTA to give MTA-modified protein. The MTA-modified protein readily reacts with FOL-modified PE-iFluor™ 660 Tandem (provided) to give the desired PE-iFluor™ 660 Tandem-antibody conjugate.

SAMPLE EXPERIMENTAL PROTOCOL

Preparation of pre-activated Antibody with Buccutite™ MTA

Reconstitute Buccutite™ MTA in DMSO at ~10 mg/mL.
Note Store unused MTA at -20 °C; it can be used for up to two freeze and thaw cycles.
Prepare target antibody (Ab) in pH = 8.5 - 9.0 buffer at a concentration above 1 mg/ml.
Add the MTA to Ab solution at the ratio of 8 - 10 µg MTA/100 µg Ab.
Mix well and react at room temperature for 60 minutes, rotating during the reaction.
Purify the reaction mixture with a desalting column to remove any unreacted MTA. Exchange the buffer to PBS or another buffer of your choice.
Collect the MTA-activated Ab. Estimate the concentration by 70% yield of the original starting amount.

Conjugate with Pre-activated PE-iFluor™ 660 Tandem

Reconstitute pre-activated PE-iFluor™ 660 Tandem in 100 µL ddH₂O to 10 mg/mL.
Note Reconstituted pre-activated PE-iFluor™ 660 Tandem is not stable and can not be stored for more than one month.
Add pre-activated PE-iFluor™ 660 Tandem directly to MTA-activated target Ab solution at the ratio of 300 µg PE-iFluor™ 660 Tandem/100 µg MTA-activated Ab.
Rotate the mixture for 1 - 2 hours at room temperature.
The Ab/PE--iFluor™ 660 Tandem conjugates are now ready to use.
Note The antibody conjugate should be stored at >0.5 mg/mL in the presence of a carrier protein (e.g., 0.1% bovine serum albumin) and 0.02-0.05% sodium azide.
Note The Ab/PE-iFluor™ 660 Tandem can be stored at 4 °C for two months.
Optional: Ab/PE-iFluor™ 660 Tandem can be further purified through size exclusion chromatography to get better performance.

Spectrum

Open in Advanced Spectrum Viewer

Spectral properties

Absorbance (nm)	566
Extinction coefficient (cm ^-1 M ^-1)	1960000
Excitation (nm)	565
Emission (nm)	695

Product Family

Name	Excitation (nm)	Emission (nm)	Extinction coefficient (cm ^-1 M ^-1)
ReadiUse™ Preactivated PE-iFluor® 647 Tandem	565	666	1960000
ReadiUse™ Preactivated PE-iFluor® 750 Tandem	565	778	1960000
ReadiUse™ Preactivated PE-iFluor® 594 Tandem	565	606	1960000
ReadiUse™ Preactivated PE-iFluor® 700 Tandem	565	708	1960000

Images

Figure 1. Our preactivated PE-iFluor® 660 Tandem was premodified with our Buccutite™ FOL (provided). Your antibody (or other proteins) is modified with our Buccutite™ MTA (provided as free sample) to give MTA-modified protein (such as antibody). The MTA-modified protein readily reacts with FOL-modified PE-iFluor® 660 Tandem (provided) to give the desired PE-iFluor® 660 Tandem-antibody conjugate in much higher yield than the SMCC chemistry. In addition our preactivated PE-iFluor® 660 Tandem reacts with MTA-modified biopolymers at much lower concentrations than the SMCC chemistry.

References

View all 50 references: Citation Explorer

One-Pot Fabrication of Pd Nanoparticles@Covalent-Organic-Framework-Derived Hollow Polyamine Spheres as a Synergistic Catalyst for Tandem Catalysis.
Authors: Yang, Xinyi and He, Yajun and Li, Liuyi and Shen, Jinni and Huang, Jianhui and Li, Lingyun and Zhuang, Zanyong and Bi, Jinhong and Yu, Yan
Journal: Chemistry (Weinheim an der Bergstrasse, Germany) (2020): 1864-1870

Analysis of Phospholipids, Lysophospholipids, and Their Linked Fatty Acyl Chains in Yellow Lupin Seeds (Lupinus luteus L.) by Liquid Chromatography and Tandem Mass Spectrometry.
Authors: Calvano, Cosima Damiana and Bianco, Mariachiara and Ventura, Giovanni and Losito, Ilario and Palmisano, Francesco and Cataldi, Tommaso R I
Journal: Molecules (Basel, Switzerland) (2020)

Detection of Phytoestrogen Metabolites in Breastfed Infants' Urine and the Corresponding Breast Milk by Liquid Chromatography-Tandem Mass Spectrometry.
Authors: Min, Jialing and Wang, Zitong and Liang, Chenglin and Li, Wenjie and Shao, Jie and Zhu, Kunrui and Zhou, Lu and Cheng, Jing and Luo, Shihong and Yu, Lehan and Wu, Yudong and Xie, Mingyong and Hu, Xiaojuan
Journal: Journal of agricultural and food chemistry (2020): 3485-3494

Identification and quantification of phospholipids in strawberry seeds and pulp (Fragaria × ananassa cv San Andreas) by liquid chromatography with electrospray ionization and tandem mass spectrometry.
Authors: Bianco, Mariachiara and Calvano, Cosima D and Huseynli, Lachin and Ventura, Giovanni and Losito, Ilario and Cataldi, Tommaso R I
Journal: Journal of mass spectrometry : JMS (2020): e4523

A monolithic nanostructured-perovskite/silicon tandem solar cell: feasibility of light management through geometry and materials selection.
Authors: Elshorbagy, Mahmoud H and López-Fraguas, Eduardo and Chaudhry, Fateh A and Sánchez-Pena, José Manuel and Vergaz, Ricardo and García-Cámara, Braulio
Journal: Scientific reports (2020): 2271

Lipid alterations in the skeletal muscle tissues of mice after weight regain by feeding a high-fat diet using nanoflow ultrahigh performance liquid chromatography-tandem mass spectrometry.
Authors: Eum, Jung Yong and Lee, Gwang Bin and Yi, Sun Shin and Kim, Il Yong and Seong, Je Kyung and Moon, Myeong Hee
Journal: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences (2020): 122022

Supercritical CO2 Extraction and Identification of Ginsenosides in Russian and North Korean Ginseng by HPLC with Tandem Mass Spectrometry.
Authors: Razgonova, Mayya and Zakharenko, Alexander and Shin, Tai-Sun and Chung, Gyuhwa and Golokhvast, Kirill
Journal: Molecules (Basel, Switzerland) (2020)

Analysis of oxidised and glycated aminophospholipids: Complete structural characterisation by C30 liquid chromatography-high resolution tandem mass spectrometry.
Authors: Colombo, Simone and Criscuolo, Angela and Zeller, Martin and Fedorova, Maria and Domingues, M Rosário and Domingues, Pedro
Journal: Free radical biology & medicine (2019): 144-155

Fast and Quantitative Phospholipidomic Analysis of SH-SY5Y Neuroblastoma Cell Cultures Using Liquid Chromatography-Tandem Mass Spectrometry and 31P Nuclear Magnetic Resonance.
Authors: Jakubec, Martin and Bariås, Espen and Kryuchkov, Fedor and Hjørnevik, Linda Veka and Halskau, Øyvind
Journal: ACS omega (2019): 21596-21603

Plasma lipid profile comparison of five different cancers by nanoflow ultrahigh performance liquid chromatography-tandem mass spectrometry.
Authors: Lee, Gwang Bin and Lee, Jong Cheol and Moon, Myeong Hee
Journal: Analytica chimica acta (2019): 117-126