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ReadiUse™ Preactivated PE-iFluor® 660 Tandem
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.
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
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 |
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
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)
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
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
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
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
Page updated on November 20, 2024
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