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250mg | ||
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ln Vitro |
1. miRNA preparation 1.1 The miRNA product is a clear or translucent dry film that is affixed to the wall of the tube. Before using, give the tube a quick centrifugation to make sure the miRNA is at the bottom. Reconstitute the miRNA in nuclease-free water to provide a stock solution with a concentration of 20 μM. To dissolve 5 nmol of miRNA, add 250 μL of nuclease-free water and vortex many times. To dissolve 20 nmol of miRNA, add 1000 μL of nuclease-free water and vortex many times. 1.2 (Preferential) To cut down on the number of freezing and thawing times, dispense the miRNA into one or more tubes (<5). 1.3 Keep the ready mother solution between -20°C and -80°C. 2. Getting cells ready 2.1 Seed cells beforehand, and after the desired cell density is obtained, carry out cell transfection. Results of transfection are influenced by cell viability prior to transfection. 3. Adaptation 3.1 Making transfection mixes A and B ready Note: We advise using a working concentration of 100 nM for miRNA inhibitors. The ideal concentrations of miRNA products for various cell types and research objectives vary based on the selected miRNA, cell line, and analytic technique. Optimization tests should be carried out utilizing various concentrations in order to ascertain the concentration that yields the optimum outcomes. The range of concentrations that is optimal is 20~500nM. In each of the six well plates: B: 121 μL serum-free medium + 4 μL PolyFast transfection reagent; A: 120 μL serum-free medium + 5 μL miRNA. The 24-well plate's individual wells: B: 24 μL serum-free medium + 1 μL PolyFast transfection reagent; A: 23.75 μL serum-free medium + 1.25 μL miRNA. The contents of each well in the 96-well plate are as follows: B: 4.8 μL serum-free medium + 0.2 μL PolyFast transfection reagent; A: 4.75 μL serum-free medium + 0.25 μL miRNA. If you use transfection reagents from a different brand, you must modify the amount added based on the particular circumstance. 3.2 Combine the weaker versions of A and B. For fifteen minutes, incubate at room temperature. 3.3 Take out the culture medium and give the cells a PBS wash. 3.4 Give the cells the mixture of miRNA-transfection reagents. Add 250 μL of the transfection mixture (A+B) and mix thoroughly after adding 1750 μL of new culture media without double antibody to each well of the 6-well plate. with each well of the 24-well plate, add 50 μL of the transfection mixture (A+B) and well mix with 450 μL of new culture media without double antibodies. Put 10 μL of the transfection mixture (A+B) and mix thoroughly after adding 90 μL of new culture media without double antibodies to each well of the 96-well plate. 3.5 Let cells sit at 37°C for one to three days. After that, transfected cells were examined.
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Molecular Weight |
5487.69
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Related CAS # |
mmu-let-7k antagomir
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Appearance |
Typically exists as solids (or liquids in special cases) at room temperature
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HS Tariff Code |
2934.99.9001
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Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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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
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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.) |
Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
1 mM | 0.1822 mL | 0.9111 mL | 1.8223 mL | |
5 mM | 0.0364 mL | 0.1822 mL | 0.3645 mL | |
10 mM | 0.0182 mL | 0.0911 mL | 0.1822 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.