ReadiUse™ Preactivated PE-iFluor® 750 Tandem
Ordering information
| Price | |
| Catalog Number | |
| Unit Size | |
| Quantity |
Additional ordering information
| Telephone | 1-800-990-8053 |
| Fax | 1-800-609-2943 |
| sales@aatbio.com | |
| Quotation | Request |
| International | See distributors |
| Shipping | Standard overnight for United States, inquire for international |
Physical properties
| Solvent | Water |
Spectral properties
| Extinction coefficient (cm -1 M -1) | 1960000 |
| Excitation (nm) | 565 |
| Emission (nm) | 778 |
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 |
Related products
| Overview |  SDSProtocol |
See also: Flow Cytometry Reagents, Fluorescence Activated Cell Sorting (FACS), PE and APC, Spectral Flow Cytometry
Extinction coefficient (cm -1 M -1) 1960000 | Excitation (nm) 565 | Emission (nm) 778 |
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 PE-iFluor® 750 Tandem is an excellent replacement for PE-Cy7® and PE-Alexa Fluor® 750 Tandem due to their similar spectral properties. ReadiUse™ Preactivated PE-iFluor® 750 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™ 750 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™ 750 Tandem (provided) to give the desired PE-iFluor™ 750 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™ 750 Tandem
- Reconstitute pre-activated PE-iFluor™ 750 Tandem in 100 µL ddH2O to 10 mg/mL.
Note Reconstituted pre-activated PE-iFluor™ 750 Tandem is not stable and can not be stored for more than one month. - Add pre-activated PE-iFluor™ 750 Tandem directly to MTA-activated target Ab solution at the ratio of 300 µg PE-iFluor™ 750 Tandem/100 µg MTA-activated Ab.
- Rotate the mixture for 1 - 2 hours at room temperature.
- The Ab/PE--iFluor™ 750 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™ 750 Tandem can be stored at 4 °C for two months. - Optional: Ab/PE-iFluor™ 750 Tandem can be further purified through size exclusion chromatography to get better performance.
Spectrum
Open in Advanced Spectrum Viewer
Spectral properties
| Extinction coefficient (cm -1 M -1) | 1960000 |
| Excitation (nm) | 565 |
| Emission (nm) | 778 |
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® 594 Tandem | 565 | 606 | 1960000 |
| ReadiUse™ Preactivated PE-iFluor® 700 Tandem | 565 | 708 | 1960000 |
| ReadiUse™ Preactivated PE-iFluor® 660 Tandem | 565 | 695 | 1960000 |
| ReadiUse™ Preactivated APC-iFluor® 750 Tandem | 651 | 793 | 700000 |
Images
Figure 1. Our preactivated PE-iFluor® 750 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® 750 Tandem (provided) to give the desired PE-iFluor® 750 Tandem-antibody conjugate in much higher yield than the SMCC chemistry. In addition our preactivated PE-iFluor® 750 Tandem reacts with MTA-modified biopolymers at much lower concentrations than the SMCC chemistry.
References
View all 46 references: Citation Explorer
Chromophore attachment to phycobiliprotein beta-subunits: phycocyanobilin:cysteine-beta84 phycobiliprotein lyase activity of CpeS-like protein from Anabaena Sp. PCC7120
Authors: Zhao KH, Su P, Li J, Tu JM, Zhou M, Bubenzer C, Scheer H.
Journal: J Biol Chem (2006): 8573
Authors: Zhao KH, Su P, Li J, Tu JM, Zhou M, Bubenzer C, Scheer H.
Journal: J Biol Chem (2006): 8573
Excitation energy transfer from phycobiliprotein to chlorophyll d in intact cells of Acaryochloris marina studied by time- and wavelength-resolved fluorescence spectroscopy
Authors: Petrasek Z, Schmitt FJ, Theiss C, Huyer J, Chen M, Larkum A, Eichler HJ, Kemnitz K, Eckert HJ.
Journal: Photochem Photobiol Sci (2005): 1016
Authors: Petrasek Z, Schmitt FJ, Theiss C, Huyer J, Chen M, Larkum A, Eichler HJ, Kemnitz K, Eckert HJ.
Journal: Photochem Photobiol Sci (2005): 1016
Single-molecule spectroscopy selectively probes donor and acceptor chromophores in the phycobiliprotein allophycocyanin
Authors: Loos D, Cotlet M, De Schryver F, Habuchi S, Hofkens J.
Journal: Biophys J (2004): 2598
Authors: Loos D, Cotlet M, De Schryver F, Habuchi S, Hofkens J.
Journal: Biophys J (2004): 2598
Isolation and characterisation of phycobiliprotein rich mutant of cyanobacterium Synechocystis sp
Authors: Prasanna R, Dhar DW, Dominic TK, Tiwari ON, Singh PK.
Journal: Acta Biol Hung (2003): 113
Authors: Prasanna R, Dhar DW, Dominic TK, Tiwari ON, Singh PK.
Journal: Acta Biol Hung (2003): 113
Evaluation of Tolypothrix germplasm for phycobiliprotein content
Authors: Prasanna R, Prasanna BM, Mohammadi SA, Singh PK.
Journal: Folia Microbiol (Praha) (2003): 59
Authors: Prasanna R, Prasanna BM, Mohammadi SA, Singh PK.
Journal: Folia Microbiol (Praha) (2003): 59
Co-ordinated expression of phycobiliprotein operons in the chromatically adapting cyanobacterium Calothrix PCC 7601: a role for RcaD and RcaG
Authors: Noubir S, Luque I, Ochoa de Alda JA, Perewoska I, T and eau de Marsac N, Cobley JG, Houmard J.
Journal: Mol Microbiol (2002): 749
Authors: Noubir S, Luque I, Ochoa de Alda JA, Perewoska I, T and eau de Marsac N, Cobley JG, Houmard J.
Journal: Mol Microbiol (2002): 749
Phycobiliprotein genes of the marine photosynthetic prokaryote Prochlorococcus: evidence for rapid evolution of genetic heterogeneity
Authors: Ting CS, Rocap G, King J, Chisholm SW.
Journal: Microbiology (2001): 3171
Authors: Ting CS, Rocap G, King J, Chisholm SW.
Journal: Microbiology (2001): 3171
Phycobiliprotein-Fab conjugates as probes for single particle fluorescence imaging
Authors: Triantafilou K, Triantafilou M, Wilson KM.
Journal: Cytometry (2000): 226
Authors: Triantafilou K, Triantafilou M, Wilson KM.
Journal: Cytometry (2000): 226
Novel activity of a phycobiliprotein lyase: both the attachment of phycocyanobilin and the isomerization to phycoviolobilin are catalyzed by the proteins PecE and PecF encoded by the phycoerythrocyanin operon
Authors: Zhao KH, Deng MG, Zheng M, Zhou M, Parbel A, Storf M, Meyer M, Strohmann B, Scheer H.
Journal: FEBS Lett (2000): 9
Authors: Zhao KH, Deng MG, Zheng M, Zhou M, Parbel A, Storf M, Meyer M, Strohmann B, Scheer H.
Journal: FEBS Lett (2000): 9
Phycobiliprotein and fluorescence immunological assay
Authors: Wu P., undefined
Journal: Sheng Li Ke Xue Jin Zhan (2000): 82
Authors: Wu P., undefined
Journal: Sheng Li Ke Xue Jin Zhan (2000): 82