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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.
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.
Ordering information
Price ()
Catalog Number2579
Unit Size
Find Distributor
Additional ordering information
Telephone1-408-733-1055
Fax1-408-733-1304
Emailsales@aatbio.com
InternationalSee distributors
ShippingStandard overnight for United States, inquire for international
Spectral properties
Absorbance (nm)566
Extinction coefficient (cm -1 M -1)1960000
Excitation (nm)566
Emission (nm)695
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
UNSPSC12171501
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OverviewpdfSDSpdfProtocol


Absorbance (nm)
566
Extinction coefficient (cm -1 M -1)
1960000
Excitation (nm)
566
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


A: ReadiUse™ Preactivated PE-iFluor® 660 Tandem1 vial (1 mg)
B: Buccutite™ MTA1 vial (100 µg)
C: Spin Desalting ColumnNot Included

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
  1. 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.
  2. Prepare target antibody (Ab) in pH = 8.5 - 9.0 buffer at a concentration above 1 mg/ml.
  3. Add the MTA to Ab solution at the ratio of 8 - 10 µg MTA/100 µg Ab.
  4. Mix well and react at room temperature for 60 minutes, rotating during the reaction.
  5. Purify the reaction mixture with a desalting column to remove any unreacted MTA. Exchange the buffer to PBS or another buffer of your choice.
  6. Collect the MTA-activated Ab. Estimate the concentration by 70% yield of the original starting amount. 

Conjugate with Pre-activated PE-iFluor™ 660 Tandem
  1. 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.
  2. 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.
  3. Rotate the mixture for 1 - 2 hours at room temperature.
  4. 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.
  5. Optional: Ab/PE-iFluor™ 660 Tandem can be further purified through size exclusion chromatography to get better performance. 

Spectrum


Open in Advanced Spectrum Viewer
spectrum

Spectral properties

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

Product family


NameExcitation (nm)Emission (nm)Extinction coefficient (cm -1 M -1)
ReadiUse™ Preactivated PE-iFluor® 647 Tandem5696661960000
ReadiUse™ Preactivated PE-iFluor® 750 Tandem566778-
ReadiUse™ Preactivated PE-iFluor® 594 Tandem5666061960000
ReadiUse™ Preactivated PE-iFluor® 700 Tandem5667081960000

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