Br-DAPI is a labeling dye of the DAPI series. DAPI is a fluorescent dye that binds strongly to DNA. DAPI binds to the A/T base pairs of the minor groove of double-stranded DNA (one DAPI molecule can occupy 3 base pair positions), thereby amplifying the fluorescence intensity approximately 20 times. DNA can be measured by looking at the intensity of the fluorescence. DAPI is frequently used to stain both live and fixed cells because it can also pass through intact cell membranes. How to store: Keep out of direct sunlight.

Physicochemical Properties

Molecular Formula	C16H14BRN5
Molecular Weight	356.219861507416
Exact Mass	355.04
CAS #	2387906-44-5
PubChem CID	162642091
Appearance	Light yellow to yellow solid powder
LogP	2.3
Hydrogen Bond Donor Count	5
Hydrogen Bond Acceptor Count	2
Rotatable Bond Count	3
Heavy Atom Count	22
Complexity	446
Defined Atom Stereocenter Count	0
SMILES	BrC1=C(C2C=CC(C(=N)N)=CC=2)NC2C=C(C(=N)N)C=CC=21
InChi Key	GSBXNZFEZFIMCS-UHFFFAOYSA-N
InChi Code	InChI=1S/C16H14BrN5/c17-13-11-6-5-10(16(20)21)7-12(11)22-14(13)8-1-3-9(4-2-8)15(18)19/h1-7,22H,(H3,18,19)(H3,20,21)
Chemical Name	3-bromo-2-(4-carbamimidoylphenyl)-1H-indole-6-carboximidamide
Synonyms	Br-DAPI
HS Tariff Code	2934.99.9001
Storage	Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: This product requires protection from light (avoid light exposure) during transportation and storage.
Shipping Condition	Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Biological Activity

ln Vitro

Usage instructions 1. Creating the functioning DAPI solution 1.1 Getting the stock solution ready To make a 1 mg/mL stock solution, dilute 1 milligram of DAPI with 1 mL of double-distilled water. It is advised to aliquot the DAPI storage solution and keep it in the dark at -20°C or -80°C. 1.2 Making a functional solution To make a 1-10 μg/mL DAPI working solution, use either PBS dilution storage solution or hot serum-free cell culture media. Note: Before using the DAPI working fluid, please make sure that its concentration is appropriate for the current circumstances. 2. Staining of suspended cells 2.1 Centrifuge the cells to collect them, then add PBS and wash twice for five minutes each time. There are 1×106 cells per milliliter. 2.2 Add 1 milliliter of DAPI working solution, then let it sit at room temperature for three to ten minutes. 2.3 After centrifuging for three to four minutes at 400 g, remove the supernatant. 2.4 Add PBS and give the cells two 5-minute washings. 2.5 Re-suspend the cells in 1 mL of PBS or serum-free media, and use a flow cytometer or fluorescence microscope to observe. 3. Adherent cell staining 3.1 Use sterile coverslips to culture adherent cells. 3.2 Aspirate the extra culture medium after removing the coverslip from the medium. 3.3 Add 100 μL of the dye working solution, give the cells a gentle shake to cover them completely, and then let them sit for three to ten minutes. 3.4 Aspirate the dye working solution, wash with culture medium two or three times for five minutes each time, and use a flow cytometer or fluorescence microscope to monitor. Storage conditions: -20°C, one year of light protection. Notes: 1. Please modify the concentration of the DAPI working solution and the incubation period in accordance with the actual circumstances. 2. Professionals may only use this product for scientific study; it may not be utilized for clinical diagnosis or treatment, nor may it be included into food or medication. 3. Please wear a lab coat and disposable gloves for your own health and safety.

References

[1]. DAPI: a DNA-specific fluorescent probe. Biotech Histochem. 1995 Sep;70(5):220-33.

Solubility Data

Solubility (In Vitro)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

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 (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *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). 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)] *Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL 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). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.8073 mL	14.0363 mL	28.0725 mL
	5 mM	0.5615 mL	2.8073 mL	5.6145 mL
	10 mM	0.2807 mL	1.4036 mL	2.8073 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.