【貨號(hào)】 BC-CL03【規(guī)格】 1ml/1.5ml
【保存】 2-8℃，12個(gè)月
【類(lèi)型】 陽(yáng)離子聚合物
【產(chǎn)品介紹】
BC Trans-mRNA是一款高性能的專(zhuān)用轉(zhuǎn)染試劑，專(zhuān)為mRNA設(shè)計(jì)，適用于在多種貼壁細(xì)胞中進(jìn)行mRNA轉(zhuǎn)染。這款試劑的獨(dú)特之處在于，它能夠直接將mRNA傳遞到細(xì)胞質(zhì)中進(jìn)行表達(dá)，從而巧妙地繞過(guò)了轉(zhuǎn)錄調(diào)控作用以及進(jìn)入細(xì)胞核的限制。因此，BC Trans-mRNA成為短期蛋白表達(dá)相關(guān)研究工作的理想選擇，為科研人員提供了便捷、高效的工具。
【應(yīng)用范圍】
BC Trans-mRNA轉(zhuǎn)染試劑展現(xiàn)出廣泛的適用性，能夠高效應(yīng)對(duì)眾多貼壁或懸浮細(xì)胞株的mRNA轉(zhuǎn)染需求。無(wú)論是在常規(guī)細(xì)胞、難以轉(zhuǎn)染的細(xì)胞、原代細(xì)胞還是干細(xì)胞中，該試劑都能實(shí)現(xiàn)較高的轉(zhuǎn)染效率，并且性能穩(wěn)定可靠。與其他轉(zhuǎn)染試劑相比，BC Trans-mRNA對(duì)血清的干擾極小、毒性低、穩(wěn)定性好、操作簡(jiǎn)便易行以及良好的重復(fù)性。
【mRNA的轉(zhuǎn)染步驟】
以24孔板為例，請(qǐng)參考表1的轉(zhuǎn)染規(guī)模調(diào)整，步驟如下:
1. 細(xì)胞接種: 每孔接種0.5~1.0×105個(gè)細(xì)胞，細(xì)胞培養(yǎng)12~24小時(shí)，使轉(zhuǎn)染時(shí)細(xì)胞密度達(dá)到60~70%融合度。
2. mRNA稀釋: 將mRNA稀釋于Opti-MEM培養(yǎng)基中，終濃度0.1mg / mL。
3. 復(fù)合物制備:
（1）取1µL的BC Trans-mRNA試劑原液加入到9µL Opti-MEM培養(yǎng)基中
（2）取4µL濃度為0.1mg / mL的mRNA加入到6µL Opti-MEM培養(yǎng)基中
（3）兩者等體積混合
（4）室溫靜置10分鐘
4. 轉(zhuǎn)染細(xì)胞：將20µL的轉(zhuǎn)染復(fù)合物均勻加入到細(xì)胞培養(yǎng)板的每個(gè)孔中，并輕輕混勻。
5. 培養(yǎng)與檢測(cè)：將細(xì)胞在37℃下繼續(xù)培養(yǎng)18~48小時(shí)。在此期間，無(wú)需更換培養(yǎng)基。
【mRNA的轉(zhuǎn)染優(yōu)化】
可通過(guò)改變細(xì)胞密度、 mRNA濃度以及BC Trans-mRNA濃度對(duì)轉(zhuǎn)染進(jìn)行優(yōu)化。保證細(xì)胞融合度在60%以上，BC Trans-mRNA (µL): mRNA (µg)可以在2:1和5:1之間調(diào)整。
表1. 不同培養(yǎng)板所需轉(zhuǎn)染試劑和mRNA的用量
| 培養(yǎng)板 | | 接種 培養(yǎng)基 | Opti-MEM培養(yǎng) 基釋后終體積 | mRNA轉(zhuǎn)染 |
| 試劑用量 | mRNA |
| 96孔板 | 0.3cm2 | 200µL | 10µL | 0.5µL | 0.2µg |
| 24孔板 | 2.0cm2 | 500µL | 20µL | 1µL | 0.4µg |
| 12孔板 | 4.0cm2 | 1mL | 40µL | 2.5µL | 1µg |
| 6孔板 | 10.0cm2 | 2mL | 100µL | 5µL | 2µg |
| 60mm | 20.0cm2 | 5mL | 200µL | 10µL | 4.0µg |
| 10cm | 60.0cm2 | 15mL | 600µL | 30µL | 12µg |
標(biāo)簽：轉(zhuǎn)染
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作者單位：東南大學(xué)
使用產(chǎn)品：Human MCP-1, TNF-α, TGF-β ELISA kit
2、Juan Zhang,Meng Cao,Wenwen Yang,etc.Inhibition of Glucose-6-Phosphate Dehydrogenase Could Enhance 1,4-Benzoquinone-Induced Oxidative Damage in K562 Cells[J].Oxid Med Cell Longev.2016:1-11.
作者單位：東南大學(xué)
使用產(chǎn)品：Human NADP, NADPH ELISA Kit
3、Qian-NanDi，Wei-XinCao，RunXu,etc.Chronic low-dose exposure of nonylphenol alters energy homeostasis in the reproductive system of female rats[J].Toxicology and Applied Pharmacology.2018,1(348):67-75.
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4、Muhammad Noman Khan,Bing Wang,Jing Wei,etc.CXCR1/2 antagonism with CXCL8/Interleukin-8 analogue CXCL8(3-72)K11R/G31P restricts lung cancer growth by inhibiting tumor cell proliferation and suppressing angiogenesis[J].Oncotarget.2015,6(25): 21315-21327.
作者單位：國(guó)立清華大學(xué)（臺(tái)灣）
使用產(chǎn)品：Human CXCL1, CXCL6, CXCL8 ELISA kit
5、Cheng Yu Wang,Xiang Dan Li,Zhi Hong Hao,etc.Insulin-like growth factor-1 improves diabetic cardiomyopathy through antioxidative and anti-inflammatory processes along with modulation of Akt/GSK-3β signaling in rats[J].Korean J Physiol Pharmacol.2016,20(6): 613-619.
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使用產(chǎn)品：Rat IL-1β, TNF-a ELISA kit
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使用產(chǎn)品：Human IL-6 ELISA kit
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8、Jian’an Bai,He Na,Xiumei Hua,etc.A retrospective study of NENs and miR-224 promotes apoptosis of BON-1 cells by targeting PCSK9 inhibition[J].Oncotarget.2017,8(4): 6929-6939.
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9、Zhan Zhang, Di Wang,Shanlei Qiao,etc.Metabolic and microbial signatures in rat hepatocellular carcinoma treated with caffeic acid and chlorogenic acid[J].Scientific reports.2017(7):1-10.
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使用產(chǎn)品：Rat LPS ELISA kit
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使用產(chǎn)品：Human IL-1β, IL-6, TNF-α, IFN-γELISA kit
11、Xiaotong Sun,Jing Wei,Yawei Tang,etc.Leptin-induced migration and angiogenesis in rheumatoid arthritis is mediated by reactive oxygen species[J].FEBS Open Bio.2017,7(12):1899-1908.
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12、Geng Tian,Linlin Cheng,Xuewei Qi,etc.Transgenic Cotton Plants Expressing Double-stranded RNAs Target HMG-CoA Reductase (HMGR) Gene Inhibits the Growth, Development and Survival of Cotton Bollworms[J].Int J Biol Sci. 2015,11(11):1296-1305.
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13、Siyuan Cui,Lili Men,Yu Li,etc.Selenoprotein S Attenuates Tumor Necrosis Factor-α-Induced Dysfunction in Endothelial Cells[J].Mediators Of Inflamm.2018:1-16.
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14、Peng-Tao Ren,Yuan Zhang,etc.Comparative investigation of the effects of specific antigen-sensitized DC-CIK and DC-CTL cells against B16 melanoma tumor cells[J].Mol Med Rep. 2017,15(4):1533-1538.
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15、Ming Ma,Lian-mei Zhao,Xing-xiao Yang,etc.p-Hydroxylcinnamaldehyde induces the differentiation of oesophageal carcinoma cells via the cAMP-RhoA-MAPK signalling pathway[J].Scientific reports.2016(6):1-13.
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16、Yucai Chen,Tianyi Yuan,Huifang Zhang,etc.Activation of Nrf2 Attenuates Pulmonary Vascular Remodeling via Inhibiting Endothelial-to-Mesenchymal Transition: an Insight from a Plant Polyphenol[J].Int J Biol Sci.2017,13(8):1067-1081.
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17、Rao Sun,Zuoxia Zhang,Yishan Lei,etc.Hippocampal activation of microglia may underlie the shared neurobiology of comorbid posttraumatic stress disorder and chronic pain[J].Mol Pain. 2016(12):1-13.
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18、Yuanyuan Zhuang,Jiabao Liu,Pei Ma,etc.Tamarixinin A Alleviates Joint Destruction of Rheumatoid Arthritis by Blockade of MAPK and NF-κB Activation[J].Front Pharmacol.2017(8):1-11.
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20、Bo Qiu,Ming Gong,Qi-Ting He,etc.Controlled Release of Interleukin-1 Receptor Antagonist from Hyaluronic Acid-Chitosan Microspheres Attenuates Interleukin-1β-Induced Inflammation and Apoptosis in Chondrocytes[J].Biomed Res Int.2016:1-12.
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22、Yuanyuan Zhang,Minhui Sun,Yuwei Han,etc.The saponin DT-13 Attenuates Tumor Necrosis Factor-α-induced Vascular Inflammation Associated with Src/NF-кB/MAPK Pathway Modulation[J].Int J Biol Sci.2015,11(8):970-981.
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23、Ying Huang,Shaofeng Wang,Chunya Zhang,etc.Experimental study of the anti-atherosclerotic effect of demethylzeylasteral[J].Exp Ther Med.2017,13(6): 2787-2792.
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24、Lin Li,Ting Xiao,Fang Li,etc.Hydrogen sulfide reduced renal tissue fibrosis by regulating autophagy in diabetic rats[J].Mol Med Rep.2017,16(2): 1715-1722.
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使用產(chǎn)品：Rat UP ELISA kit
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27、Na Yang,Run-bin Sun,Xing-long Chen,etc.In vitro assessment of the glucose-lowering effects of berberrubine-9-O-β-D-glucuronide,an active metabolite of berberrubine[J].Acta Pharmacol Sin.2017,38(3): 351-361.
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30、Yu Li,Ning Hao,Suping Zou,etc.Immune Regulation of RAW264.7 Cells In Vitro by Flavonoids from Astragalus complanatus via Activating the NF-κB Signalling Pathway[J].Journal of Immunology Research.2018:1-9.
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32、JieChao,YanLv,JinChen,etc.SiO2-induced release of sVEGFRs from pulmonary macrophages[J].Respiratory Physiology & Neurobiology,2018(247):1-8.
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使用產(chǎn)品：Human sVEGFR ELISA Kit
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使用產(chǎn)品：Mouse IL-1β, IL-18, TNF-α ELISA Kit
34、Zhengfeng Wu,Tianhong Gao,Ronglign Zhong,etc.Antihyperlipidaemic effect of triterpenic acid-enriched fraction from Cyclocarya paliurus leaves in hyperlipidaemic rats[J].Pharmaceutical Biology,2017,1(55):712-721.
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使用產(chǎn)品：Rat apoB48 ELISA Kit
35、Rao Sun,Zhibin Zhao,Jiying Feng,etc.Glucocorticoid-Potentiated Spinal Microglia Activation Contributes to Preoperative Anxiety-Induced Postoperative Hyperalgesia[J].Mol Neurobiol.2017,54(6):4316-4328.
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使用產(chǎn)品： Rat CS, IL-1β, TNF-α ELISA Kit
36、Guoxiang You, Peifang Wang, Jun Hou,etc.Insights into the short-term effects of CeO2 nanoparticles on sludge dewatering and related mechanism[J].Water Research.2017,04(011):93-113.
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使用產(chǎn)品：Plant AHL ELISA Kit
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作者單位：江南大學(xué)
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