Br-DAPI

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.

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

C16H14BRN5

356.22

355.04

2387906-44-5

Powder

C1=CC(=CC=C1C2=C(C3=C(N2)C=C(C=C3)C(=N)N)Br)C(=N)N

GSBXNZFEZFIMCS-UHFFFAOYSA-N

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)

3-bromo-2-(4-carbamimidoylphenyl)-1H-indole-6-carboximidamide

Br-DAPI

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.

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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).

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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)

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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).

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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

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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.

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 (Please use freshly prepared in vivo formulations for optimal results.)