耗材描述
mmu-miR-12178-5p mimic
产品活性:mmu-miR-12178-5p mimic 是一种化学合成的miRNA模拟物,能模拟内源性miRNA,上调miRNA活性,用于功能获得性(gain-of-function)研究。
产品来源: https://www.medchemexpress.cn/mmu-mir-12178-5p-mimic.html
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 (
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. 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 mimics; 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 mimics; 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 mimics; B: 4.8 μL serum-free medium + 0.2 μL PolyFast Transfection Reagent (HY-K1014).
Note: The recommended working concentration is 50nM for miRNA mimics. 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 10 to 200nM.
If other transfection reagents are used, the amount of transfection reagent needs to be adjusted according to the specific situation.
3.2 Mix A and B 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 | Diosgenin | Aurintricarboxylic acid | MicroRNA Mimic Negative Control | RGLS4326 | PIN1 inhibitor API-1 | LIN28 inhibitor LI71 | Enoxacin hydrate | MicroRNA Antagomir Negative Control | Targapremir-210 | MicroRNA Inhibitor Negative Control | Pseudoprotodioscin | SID 3712249 | hsa-miR-23a-3p antagomir | hsa-miR-124-3p inhibitor | mmu-miR-155-5p mimic | Cobomarsen | hsa-miR-138-5p agomir | hsa-miR-138-5p antagomir | hsa-miR-146a-5p mimic | hsa-miR-17-5p mimic | hsa-miR-21-5p inhibitor | hsa-miR-21-5p mimic | hsa-miR-26b-5p mimic | hsa-miR-27b-3p mimic | hsa-miR-30c-5p inhibitor | Lin28-let-7a antagonist 1 | MIR96-IN-1 | hsa-miR-106a-3p inhibitor
品牌介绍: MCE (MedChemExpress) 拥有数百种全球独家化合物,我们致力于为全球科研客户提供最新最全的高品质小分子活性化合物;10,000 多种高选择性抑制剂、激动剂涉及各热门信号通路及疾病领域;设有专业的实验中心和严格的质控、验证体系;提供 LC/MS、NMR、HPLC、手性分析、元素分析等各项质检报告,确保产品的高纯度、高品质;产品的生物活性多经各国客户实验验证;Nature, Cell, Science 等多种顶级期刊及制药专利收录了MCE客户的科研成果;专业团队跟踪最新的制药及生命科学研究进展,为您提供全球最新的活性化合物;与世界各大制药公司及知名科研机构建立了长期的合作。