MCE 的所有产品仅用作科学研究或药证申报,我们不为任何个人用途提供产品和服务。
CAS No. :
MCE 国际站:hsa-miR-494-5p inhibitor
产品活性:hsa-miR-494-5p inhibitor 是一种全链经过甲氧基修饰的成熟miRNA的互补单链。miRNA inhibitor通过特异性结合成熟miRNA,阻止miRNA与其靶基因的互补配对,从而抑制miRNA的功能,可用于miRNA 功能缺失性(loss-of-function)研究。
研究领域:Epigenetics
作用靶点:MicroRNA
In Vitro:
1. miRNA Resuspension
1.1 Briefly centrifuge the tube to ensure that the dried miRNA is at the bottom of the tube.
1.2 Resuspend the miRNA using nuclease free water to generate 20 μM stock solution.
For 5 nmol miRNA: add 250 μL nuclease free water.
For 20 nmol miRNA: add 1000 μL nuclease free water.
1.3 Aliquot miRNAs into one or more tubes to limit the number of freeze-thaw cycles (<5).
1.4 Store at or below -20°C or -80°C in a non-frost-free freezer until use.
2. Prepare cells
2.1 Inoculate cells in advance for cell transfection.
Note: The viability and general health of cells prior to transfection significantly affect transfection result.
3. Transfection
3.1 Prepare transfection mix A and B.
For per well of a 6-well plate: A: 120 μL serum-free medium + 5 μL miRNA inhibitors; B: 121 μL serum-free medium + 4 μL PolyFast Transfection Reagent (HY-K1014).
For per well of a 24-well plate: A: 23.75 μL serum-free medium + 1.25 μL miRNA inhibitors; B: 24 μL serum-free medium + 1 μL PolyFast Transfection Reagent (HY-K1014).
For per well of a 96-well plate: A: 4.75 μL serum-free medium + 0.25 μL miRNA inhibitors; B: 4.8 μL serum-free medium + 0.2 μL PolyFast Transfection Reagent (HY-K1014).
Note: The recommended working concentration is 100nM for miRNA inhibitors. miRNA function can vary greatly, depending on the miRNA, the cell line, and the chosen analysis method. To determine the concentration that provides optimal results, optimization experiments using varying mimic/inhibitor concentrations should be performed. The optimized range suggests changing the miRNA concentration in the range of 20 to 500nM.
If other transfection reagents are used, the amount of transfection reagent needs to be adjusted according to the specific situation.
3.2 Mix the diluted PolyFast Transfection Reagent and miRNA gently. Incubate at room temperature for 15 minutes.
3.3 Remove culture medium from cells, wash with PBS.
3.4 Add transfection mix (A+B) to cells.
For per well of a 6-well plate: add 1750 μL fresh medium without Pen/Strep, then add 250 μL of the transfection mix (A+B) to the well, and mix well.
For per well of a 24-well plate: add 450 μL fresh medium without Pen/Strep, then add 50 μL of the transfection mix (A+B) to the well, and mix well.
For per well of a 96-well plate: add 90 μL fresh medium without Pen/Strep, then add 10 μL of the transfection mix (A+B) to the well, and mix well.
3.5 Incubate cells for 1–3 days at 37°C. Then, analyze transfected cells. The medium can be replaced with fresh serum-containing medium after 6 hours if necessary.
相关产品:Camptothecin | Cl-amidine hydrochloride | Aurintricarboxylic acid | Diosgenin | Enoxacin hydrate | LIN28 inhibitor LI71 | PIN1 inhibitor API-1 | RGLS4326 | Targapremir-210 | Pseudoprotodioscin | SID 3712249 | MicroRNA Mimic Negative Control | Lin28-let-7a antagonist 1 | MIR96-IN-1 | Gypenoside LI | hsa-miR-155-5p mimic | hsa-miR-21-5p mimic | hsa-miR-26b-5p mimic | hsa-miR-29a-3p inhibitor | hsa-miR-451a inhibitor | hsa-miR-451a mimic | Miravirsen | MTL-CEBPA | TGP-377/421 | Cobomarsen | Enoxacin-d8 hydrochloride | hsa-miR-100-5p mimic
热门产品线:重组蛋白 | 化合物库 | 天然产物 | 荧光染料 | PROTAC | 同位素标记物
Trending products:Recombinant Proteins | Bioactive Screening Libraries | Natural Products | Dye Reagents | PROTAC | Isotope-Labeled Compounds