BD Pharmingen™ DRAQ7™ 564904

Preparation And Storage: Preparation And Storage Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Recommended Assay Procedures: Recommended Assay Procedures Staining of Live Cells for Viability Analysis by Flow Cytometry 1. Obtain a single cell suspension. 2. Resuspend cells in 1× Dulbecco's Phosphate Buffered Saline (DPBS) or other azide-free buffer containing 1-3 μM DRAQ7™. a. The optimal concentration of DRAQ7™ for viability analysis may vary by cell type. We recommend titrating the reagent for your cell type of interest in early experiments. b. Additionally, apoptotic cells may stain with variable amounts of DRAQ7™. We recommend co-staining with, eg, BD Pharmingen™ FITC  Annexin V (Cat. No. 556419) if further analysis of apoptotic cells is desired. 3. Incubate 5 minutes at room temperature. No wash is necessary prior to analysis. 4. Proceed to analysis by flow cytometry. Staining of Fixed Cells for DNA Content Analysis by Flow Cytometry 1. Obtain a single cell suspension. 2. Treat cells on ice for 30 minutes with 70-80% ice-cold ethanol. a. Ethanol fixation typically provides the most resolved histograms. However, this reagent has also been successfully used for DNA content analysis with the BD Pharmingen™  Transcription Factor Buffer Set (Cat. No. 562574/562725) or BD Cytofix™ Fixation Buffer (Cat. No. 554655) and BD Phosflow™ Perm III (Cat. No. 558050) protocol. 3. Wash cells once with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656). 4. Dilute DRAQ7™ to 20 μM in 1× DPBS or other azide-free buffer immediately prior to use. 5. Stain cells for 5-15 minutes at a cell density of 0.5E6 cells/mL or less. No further wash is necessary prior to analysis. a. The optimal cell density and concentration of DRAQ7™ for DNA content analysis may vary by cell type. Assay conditions should be optimized in early experiments for best results. 6. Proceed to analysis by flow cytometry. Immunofluorescent Staining of Fixed Cells for Nuclear Visualization 1. Fix and permeabilize cells as desired. 2. Dilute DRAQ7™ solution to 5-20 μM in 1× DPBS or other azide-free buffer immediately prior to use. 3. Add DRAQ7™ solution to each sample at least 5 minutes before analysis. 4. Proceed to imaging. We recommend using a 715LP or longer wavelength filter, though the dye is well-detected in filters typically used to detect Alexa Fluor® 647 (eg, 660/20 or 692/40). Note that dsDNA-bound dye will fluoresce brightly in the nucleus and unbound dye may fluoresce dimly in the cytoplasm, allowing segmentation of the cytoplasmic and nuclear compartments. Note: This reagent has been developed and certified for the Bioimaging application. However, a routine Bioimaging test is not performed on every lot. Warning: DRAQ7™ contains < 1% 1,5-BIS{[2-(DIMETHYLAMINO)ETHYL]AMINO}-4,8-DIHYDROXYANTHRACENE-9,10-DIONE Hazard statements Causes skin irritation. Causes serious eye irritation. May cause respiratory irritation. Precautionary statements Wear protective gloves/protective clothing/eye protection/face protection. Wash thoroughly after handling. IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing. If eye irritation persists: Get medical advice/attention. Take off contaminated clothing and wash before reuse. Call a POISON CENTER or doctor/physician if you feel unwell. If skin irritation occurs: Get medical advice/attention. IF ON SKIN: Wash with plenty of water Avoid breathing dust/fume/gas/mist/vapors/spray. Use only outdoors or in a well-ventilated area. IF INHALED: Remove person to fresh air and keep comfortable for breathing. Store in a well-ventilated place. Keep container tightly closed.  Store locked up. Dispose of contents/container to an appropriate treatment and disposal facility in accordance with applicable laws and regulations, and product characteristics at time of disposal.
Product Notices: Product Notices DRAQ7™ is a registered trademark of BioStatus Ltd. FlowJo is a trademark of Tree Star Inc. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR. mTESR™1 is a trademark of StemCell Technologies. This antibody has been developed and certified for the bioimaging application. However, a routine bioimaging test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.