Flow cytometry protocols handbook
Protocols that fit your needs in flow cytometry ranging from sample preparation to numerous cell stimulation conditions, staining, immunophenotyping, and data analysis strategies.
5 | 594/633 | 660/20 | 650 | 671 | (in buffer) 3 (in antifade) 5 | microscopy, flow cytometry, fluorescence microplate reader |
Invitrogen Alexa Fluor 647 dye is a bright, far-red–fluorescent dye with excitation ideally suited for the 594 nm or 633 nm laser lines. For stable signal generation in imaging and flow cytometry, Alexa Fluor 647 dye is pH-insensitive over a wide molar range. Probes with high fluorescence quantum yield and high photostability allow detection of low-abundance biological structures with great sensitivity. Alexa Fluor 647 dye molecules can be attached to proteins at high molar ratios without significant self-quenching, leading to brighter conjugates and more sensitive detection.
We offer Alexa Fluor 647 dye conjugated to a variety of antibodies, peptides, proteins, tracers, and amplification substrates optimized for cellular labeling and detection. In addition, reactive dye forms and protein labeling kits are available to allow you to generate your own antibody conjugates or probes.
Protein Labeling Reagents Selection Guide (NHS ester, maleimide, etc.) Search Alexa Fluor 647 secondary antibodies
Figure 1. Flow cytometry histogram of cells stained with (right) and without (left) anti-CD4 biotin and Alexa Fluor 647. Human mononuclear cells were stained with mouse anti human CD4 biotin, washed, then stained with streptavidin, Alexa Fluor 647 conjugate and analyzed by flow cytometry using 633 nm excitation.
Figure 2. HeLa cells expressing CellLight Mitochondria-GFP were fixed and permeabilized. Tubulin was probed with Mouse Anti-beta-3 Tubulin Antibody and Alexa Fluor 555 Goat Anti-Mouse Secondary Antibody. Actin was stained with Alexa Fluor 647 Phalloidin and nuclei labeled with NucBlue Fixed Cell ReadyProbes Reagent (DAPI). Cells were mounted in ProLong Glass Antifade Mountant and imaged on an EVOS FL Auto2 microscope using a 20X objective.
Figure 3. HeLa cells were fixed and permeabilized with Image-iT Fixation/Permeabilization Kit. Cells were incubated with anti-Prohibitin antibody and labeled with Alexa Fluor 647 Tyramide SuperBoost Kit - Goat anti-Rabbit IgG. For Beta-Catenin detection cells were incubated with anti-Beta-Catenin and labeled with Alexa Fluor 488 Tyramide SuperBoost Kit - Goat anti-Mouse IgG. Nucleus was labeled with NucBlue Fixed Cell ReadyProbes Reagent (DAPI). Images were taken on a confocal microscope.
Protocols that fit your needs in flow cytometry ranging from sample preparation to numerous cell stimulation conditions, staining, immunophenotyping, and data analysis strategies.
A handy reference poster featuring the broad range of our available dyes and labeling reagents.
Protocols that fit your needs in imaging ranging from sample and assay preparation to staining, labeling, and data analysis strategies.
A tool for selecting the optimal fluorescent dyes for your Invitrogen EVOS cell imaging systems.
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Not for resale. Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.
Brilliant Ultra Violet™ and Brilliant Violet™ are trademarks or registered trademarks of Becton, Dickinson and Company or its affiliates, and are used under license.
For Research Use Only. Not for use in diagnostic procedures.