For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:
FC-FP
Product Information
Product Usage Information
1. Prepare the following reagents with reverse osmosis deionized (RODI) or equivalent grade water:
a. 1X PBS (azide- and protein/serum-free)
b. Incubation Buffer: Dissolve 0.5 g Bovine Serum Albumin (BSA) (#9998) in 100 ml 1X PBS. Store at 4°C.
2. Remove Ghost Dye™ from -20°C and bring to room temperature.
3. Collect cells by centrifugation and aspirate supernatant.
4. Wash cells by centrifugation in excess 1X PBS. Repeat if necessary.
5. Resuspend cells to a concentration of 1-10 x 106/mL in 1X PBS.
6. Centrifuge the Ghost Dye™ before opening then add 1 uL for each 1 mL of cell suspension and vortex immediately.
7. Incubate for 30 minutes at 4°C protected from light.
8. Wash by centrifugation in excess incubation buffer. Discard supernatant. Repeat.
9. Cells can then be fixed, permeabilized, and immunostained based upon experimental design and recommended protocols.
10. Exclude cells with high Ghost Dye™ fluorescence from analysis. These were non-viable cells at the time of fixation. See details below for excitation and emission specifications.
Ghost Dye™ Red 780 Fixable Viability Dye is excited by the red (633 nm) laser line and has a peak emission of 780 nm that can be detected using a 780/60 band pass filter commonly used for detection of APC-Cy7®
Storage
Store at -20°C desiccated and protected from light. This product is stable for 12 months. Aliquot to avoid excessive freeze-thaw cycles.
Specificity / Sensitivity
Species Reactivity:
All Species Expected
Product Description
Ghost Dye™ Red 780 Fixable Viability Dye is used to discriminate viable from non-viable mammalian cells in flow cytometry applications. Ghost Dye™ Red 780 Fixable Viability Dye irreversibly binds free amines available on the cell surface as well as intracellular free amines exposed in cells with compromised cell membranes. Non-viable cells with loss of membrane integrity will be labeled with significantly more dye than healthy cells in the same sample. The cells may then be fixed, and the degree of labeling will be preserved through fixation, permeabilization, and antibody incubation steps. Cells that exhibit increased fluorescence intensity were non-viable at the time of fixation and can be excluded from analysis.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Applications Key
FC-FP: Flow Cytometry (Fixed/Permeabilized)
Cross-Reactivity Key
H: humanM: mouseR: ratHm: hamsterMk: monkeyVir: virusMi: minkC: chickenDm: D. melanogasterX: XenopusZ: zebrafishB: bovineDg: dogPg: pigSc: S. cerevisiaeCe: C. elegansHr: horseGP: Guinea PigRab: rabbitAll: all species expected
Trademarks and Patents
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Cy and CyDye are registered trademarks of GE Healthcare.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.
Limited Uses
Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following termsapply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are inaddition to, or different from, those contained herein, unless separately accepted in writing by a legally authorizedrepresentative of CST, are rejected and are of no force or effect.
Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensedby the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnosticor therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CSTare provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic,prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose,requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make availableany Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture anycommercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlyingstructure or technology of the Products, or use the Products for the purpose of developing any products or services that wouldcompete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent orcopyright notices or markings, (d) use the Products solely in accordance withCST Product Terms of Sale and any applicabledocumentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products orservices used by Customer in connection with the Products.
