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| 100mg | ||
| 250mg | ||
| 500mg | ||
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| ln Vitro |
Mammalian cell full culture medium staining technique [2]: (1) Cell plating: Break down and separate the cells, then resuspend them in complete culture medium at a concentration of 2-4 x 105/ml. NOTE: By soaking 1-2 ml of dye for 4 cm2, attached cultured cells (such as coverslips or chambers) can be stained in situ. (2) To prepare the staining solution, add 4 µl of the 5 mM dye stock solution to each milliliter of culture medium, resulting in a final concentration of 20 µM. Note: Nuclear staining often does not surpass 30 µM and can be observed at 2.5–5 µM. (3) For fluorescent staining, incubate for five to fifteen minutes at 37°C. It should not happen to stain too much. (4) Optional cleaning: centrifuge for three to five minutes at 800 g and 37°C. Get rid of the supernatant and resuspend the cells in full media with 10 mM HEPES (HY-D0857) at a volume of 4 × 105/ml. (5) Flow cytometry: Employ standard pulse analysis for dual identification, and make use of the proper software to assess parameters. (6) Laser scanning microscope: Take fluorescence pictures with a 695 nm filter. Method for staining fixed cells [2]: (1) Fixed cells: fix for 30 minutes in PBS containing 4% paraformaldehyde, then resuspend in buffer (like PBS). (2) Fluorescent staining: Use dye concentrations and culture conditions that are comparable to those of living cells.
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| References |
[1]. Smith PJ, et al. A novel cell permeant and far red-fluorescing DNA probe, DRAQ5, for blood cell discrimination by flow cytometry. J Immunol Methods. 1999 Oct 29;229(1-2):131-9.
[2]. Smith PJ, et al. DRAQ5 labeling of nuclear DNA in live and fixed cells. Curr Protoc Cytom. 2004 May;Chapter 7:Unit 7.25. |
| Molecular Formula |
C20H24N4O4
|
|---|---|
| Molecular Weight |
384.43
|
| CAS # |
254098-36-7
|
| SMILES |
C1(O)=C2C(C(=O)C3=C(C2=O)C(NCCNC)=CC=C3O)=C(NCCNC)C=C1
|
| Synonyms |
DRAQ5
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
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| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.6013 mL | 13.0063 mL | 26.0125 mL | |
| 5 mM | 0.5203 mL | 2.6013 mL | 5.2025 mL | |
| 10 mM | 0.2601 mL | 1.3006 mL | 2.6013 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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