ReadiUse™ Preactivated PE
Ordering information
Price | |
Catalog Number | |
Unit Size | |
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Additional ordering information
Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
International | See distributors |
Bulk request | Inquire |
Custom size | Inquire |
Shipping | Standard overnight for United States, inquire for international |
Physical properties
Solvent | Water |
Spectral properties
Correction Factor (280 nm) | 0.175 |
Extinction coefficient (cm -1 M -1) | 1960000 |
Excitation (nm) | 565 |
Emission (nm) | 574 |
Quantum yield | 0.82 |
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 | ![]() ![]() |
See also: Buccutite™ Crosslinkers and Kits, PE and APC
Correction Factor (280 nm) 0.175 | Extinction coefficient (cm -1 M -1) 1960000 | Excitation (nm) 565 | Emission (nm) 574 | Quantum yield 0.82 |
R-Phycoerythrin (PE) is isolated from red algae. Its primary absorption peak is at 565 nm with secondary peaks at 496 and 545 nm. AAT Bioquest offers this preactivated PE to facilitate the PE conjugations to antibodies and other proteins such as streptavidin and other secondary reagents. Our preactivated PE is ready to conjugate, giving much higher yield than the conventionally tedious SMCC-based conjugation chemistry. In addition, our preactivated PE is conjugated to a protein via its amino group that is abundant in proteins while SMCC chemistry targets the thiol group that has to be regenerated by the reduction of antibodies.
Components
Example protocol
AT A GLANCE
Important PE 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 (provided) to give the desired PE-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
- Reconstitute pre-activated PE in 100 µL ddH2O to 10 mg/mL.
Note Reconstituted pre-activated PE is not stable and can not be stored for more than one month. - Add pre-activated PE directly to MTA-activated target Ab solution at the ratio of 300 µg PE/100 µg MTA-activated Ab.
- Rotate the mixture for 1 - 2 hours at room temperature.
- The Ab/PE 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 can be stored at 4 °C for two months. - Optional: Ab/PE can be further purified through size exclusion chromatography to get better performance.
Spectrum
Open in Advanced Spectrum Viewer
![spectrum](/_next/image?url=https%3A%2F%2Fimages.aatbio.com%2Fspectra%2Fpe_phycoerythrin.png&w=2048&q=50)
![spectrum](/_next/image?url=https%3A%2F%2Fimages.aatbio.com%2Fspectra%2Fpe_phycoerythrin.png&w=2048&q=50)
Spectral properties
Correction Factor (280 nm) | 0.175 |
Extinction coefficient (cm -1 M -1) | 1960000 |
Excitation (nm) | 565 |
Emission (nm) | 574 |
Quantum yield | 0.82 |
Product Family
Name | Excitation (nm) | Emission (nm) | Extinction coefficient (cm -1 M -1) | Correction Factor (280 nm) |
ReadiUse™ Preactivated APC | 651 | 660 | 730000 | 0.195 |
ReadiUse™ Preactivated PerCP | 477 | 678 | 406000 | 0.22 |
Images
![Our preactivated PE 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 (provided) to give the desired PE-antibody conjugate in much higher yield than the SMCC chemistry. In addition our preactivated PE reacts with MTA-modified biopolymers at much lower concentrations than the SMCC chemistry.](/_next/image?url=https%3A%2F%2Fimages.aatbio.com%2Fproducts%2Ffigures-and-data%2Freadiuse-preactivated-pe%2Ffigure-for-readiuse-preactivated-pe_XCfEE.jpg&w=3840&q=75)
Figure 1. Our preactivated PE 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 (provided) to give the desired PE-antibody conjugate in much higher yield than the SMCC chemistry. In addition our preactivated PE reacts with MTA-modified biopolymers at much lower concentrations than the SMCC chemistry.
![Top) Spectral pattern was generated using a 4-laser spectral cytometer. Spatially offset lasers (355 nm, 405 nm, 488 nm, and 640 nm) were used to create four distinct emission profiles, then, when combined, yielded the overall spectral signature. Bottom) Flow cytometry analysis of PBMC stained with PE anti-human CD4 *SK3* conjugate. The fluorescence signal was monitored using an Aurora spectral flow cytometer in the PE specific B6-A channel.](/_next/image?url=https%3A%2F%2Fimages.aatbio.com%2Fproducts%2Ffigures-and-data%2Freadiuse-preactivated-pe%2Ffigure-for-readiuse-preactivated-pe_j3ejW.png&w=3840&q=75)
Figure 2. Top) Spectral pattern was generated using a 4-laser spectral cytometer. Spatially offset lasers (355 nm, 405 nm, 488 nm, and 640 nm) were used to create four distinct emission profiles, then, when combined, yielded the overall spectral signature. Bottom) Flow cytometry analysis of PBMC stained with PE anti-human CD4 *SK3* conjugate. The fluorescence signal was monitored using an Aurora spectral flow cytometer in the PE specific B6-A channel.
References
View all 96 references: Citation Explorer
Imaging Flow Cytometry for Phylogenetic and MorphologicallyBased Functional Group Clustering of a Natural Phytoplankton Community over 1 Year in an Urban Pond.
Authors: Dunker, Susanne
Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology (2020)
Authors: Dunker, Susanne
Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology (2020)
In Vivo Photodynamic Therapy With a Lipophilic Zinc(II) Phthalocyanine Inhibits Colorectal Cancer and Induces a Th1/CD8 Antitumor Immune Response.
Authors: Chiarante, Nicolás and Duhalde Vega, Maite and Valli, Federico and Zotta, Elsa and Daghero, Hellen and Basika, Tatiana and Bollati-Fogolin, Mariela and García Vior, María C and Marino, Julieta and Roguin, Leonor P
Journal: Lasers in surgery and medicine (2020)
Authors: Chiarante, Nicolás and Duhalde Vega, Maite and Valli, Federico and Zotta, Elsa and Daghero, Hellen and Basika, Tatiana and Bollati-Fogolin, Mariela and García Vior, María C and Marino, Julieta and Roguin, Leonor P
Journal: Lasers in surgery and medicine (2020)
Purification of Live Stem-Cell-Derived Islet Lineage Intermediates.
Authors: Rasouli, Niloofar and Melton, Douglas A and Alvarez-Dominguez, Juan R
Journal: Current protocols in stem cell biology (2020): e111
Authors: Rasouli, Niloofar and Melton, Douglas A and Alvarez-Dominguez, Juan R
Journal: Current protocols in stem cell biology (2020): e111
Application Settings versus Fixed Photomultiplier Tube Voltages for Optimal Cytometer Stability over Time, and Effect on Reusing a Compensation Matrix.
Authors: Andersen, Morten Nørgaard and Skovbo, Anni and Petersen, Charlotte Christie and Hokland, Marianne
Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology (2020)
Authors: Andersen, Morten Nørgaard and Skovbo, Anni and Petersen, Charlotte Christie and Hokland, Marianne
Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology (2020)
Improved Flow Cytometric Light Scatter Detection of Submicron-Sized Particles by Reduction of Optical Background Signals.
Authors: Arkesteijn, Ger J A and Lozano-Andrés, Estefanía and Libregts, Sten F W M and Wauben, Marca H M
Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology (2020)
Authors: Arkesteijn, Ger J A and Lozano-Andrés, Estefanía and Libregts, Sten F W M and Wauben, Marca H M
Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology (2020)
Yak (Bos grunniens) bones collagen-derived peptides stimulate osteoblastic proliferation and differentiation via the activation of Wnt/β-catenin signaling pathway.
Authors: Ye, Mengliang and Zhang, Chunhui and Zhu, Lingyu and Jia, Wei and Shen, Qingshan
Journal: Journal of the science of food and agriculture (2020): 2600-2609
Authors: Ye, Mengliang and Zhang, Chunhui and Zhu, Lingyu and Jia, Wei and Shen, Qingshan
Journal: Journal of the science of food and agriculture (2020): 2600-2609
Hematologist-Level Classification of Mature B-Cell Neoplasm Using Deep Learning on Multiparameter Flow Cytometry Data.
Authors: Zhao, Max and Mallesh, Nanditha and Höllein, Alexander and Schabath, Richard and Haferlach, Claudia and Haferlach, Torsten and Elsner, Franz and Lüling, Hannes and Krawitz, Peter and Kern, Wolfgang
Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology (2020)
Authors: Zhao, Max and Mallesh, Nanditha and Höllein, Alexander and Schabath, Richard and Haferlach, Claudia and Haferlach, Torsten and Elsner, Franz and Lüling, Hannes and Krawitz, Peter and Kern, Wolfgang
Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology (2020)
Comprehensive Phenotyping of Human PB NK Cells by Flow Cytometry.
Authors: Del Zotto, Genny and Antonini, Francesca and Pesce, Silvia and Moretta, Francesca and Moretta, Lorenzo and Marcenaro, Emanuela
Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology (2020)
Authors: Del Zotto, Genny and Antonini, Francesca and Pesce, Silvia and Moretta, Francesca and Moretta, Lorenzo and Marcenaro, Emanuela
Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology (2020)
In Vitro Evaluation of Toxicant Influences on the Immune System.
Authors: Kasten-Jolly, Jane and Lawrence, David A
Journal: Current protocols in toxicology (2020): e95
Authors: Kasten-Jolly, Jane and Lawrence, David A
Journal: Current protocols in toxicology (2020): e95
Procedures for Flow Cytometry-Based Sorting of Unfixed Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Infected Cells and Other Infectious Agents.
Authors: Reifel, Kristen M and Swan, Brandon K and Jellison, Evan R and Ambrozak, David and Baijer, Jan and Nguyen, Richard and Monard, Simon and Lyon, Geoffrey and Fontes, Benjamin and Perfetto, Stephen P
Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology (2020)
Authors: Reifel, Kristen M and Swan, Brandon K and Jellison, Evan R and Ambrozak, David and Baijer, Jan and Nguyen, Richard and Monard, Simon and Lyon, Geoffrey and Fontes, Benjamin and Perfetto, Stephen P
Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology (2020)
Application notes
A Meta-Analysis of Common Calcium Indicators
A New Protein Crosslinking Method for Labeling and Modifying Antibodies
A Novel Fluorescent Probe for Imaging and Detecting Hydroxyl Radical in Living Cells
A Novel NO Wash Probeniceid-Free Calcium Assay for Functional Analysis of GPCR and Calcium Channel Targets
Abbreviation of Common Chemical Compounds Related to Peptides
A New Protein Crosslinking Method for Labeling and Modifying Antibodies
A Novel Fluorescent Probe for Imaging and Detecting Hydroxyl Radical in Living Cells
A Novel NO Wash Probeniceid-Free Calcium Assay for Functional Analysis of GPCR and Calcium Channel Targets
Abbreviation of Common Chemical Compounds Related to Peptides