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Namipo 轉染試劑
發布日期:2017-09-02 12:38:35 瀏覽次數 :
導讀
NamipoTM 轉染試劑Nanoscale micro poly Transfection Reagent方便 無毒 高效產品簡介NamipoTM是一種高分子納米微聚球結構轉染試劑

广东快乐十分选一数投:NamipoTM 轉染試劑
Nanoscale micro poly  Transfection  Reagent
 方便   無毒   高效
 

產品簡介

NamipoTM是一種高分子納米微聚球結構轉染試劑,具有生物相容性,可生物降解。特別適用于RNAi的體外轉染,也可以轉染高純度的水溶質粒。Namipo轉染時不需要無血清培養,抗生素無影響,不需要與培養基共同孵育,不需要換液,且具備核酸緩釋和?;さ墓δ?。


產品優勢

效率高一點

广东快乐十分直播开奖 www.netjwr.com.cn Namipo相對于傳統的脂質體試劑,表現出更好的轉染效率,克服了原代細胞的RNAi轉染和部分原代細胞的質粒轉染。
  \       \
        siRNA轉染原代神經元                                                               質粒轉染原代成纖維

毒性小一點

轉染無需換液,相對于傳統質粒體,轉染后24h細胞密度至少高出20%。使用Namipo轉染RNAi可獲得更高的干擾效率。
\       \
      SiRNA轉染RAW264.7細胞狀態                                       電鏡下的Namipo
 


價格低一點

一樣的使用比例下,進口試劑是我們價格的一到三倍,超高的性價比也是您選擇Namipo一個合適理由。

Catalog #

TSBBR001

TSBBR003

TSBBR0015

TSBBR0075

Size

0.3 mL

1mL

1.5mL

5X1.5mL

單價 (CNY)

800

2200

3200

15000


使用方法

 

  • siRNA 轉染(RNAi)

RNAi Transfection
 

A.將siRNA溶于RNase-Free water ,震蕩混勻配置成20μM母液,配置方法見附表一。

B.使用前請用振蕩器輕微震蕩5s混勻,如有少量沉淀屬于正常現象,本品呈透明或乳白色均不影響產品品質。

C.根據使用量(參考附表三)將Namipo(μl)與siRNA母液(μl) 1:1混合,移液器輕輕吹打10次以上充分混勻,室溫或4℃孵育10分鐘。

D.按照使用量將Namipo-SiRNA復合物加入培養基,吹打5-10次混勻,或8字搖勻。轉染前后培養基無需特殊處理,24小時內建議不要換液。

E.轉染后檢測,若所轉染的siRNA帶有熒光標記,可在轉染12小時后直接觀察熒光,觀察轉染效率。若siRNA不帶熒光,轉染24-48小時可通過qPCR檢測靶基因的表達水平,由于siRNA本身就存在效率問題,為了方便判斷轉染是否成功,最好設置一組GAPDH的siRNA陽性對照(NamipoTM  轉染試劑試劑盒會贈送0.5nmol Fam-NC綠色熒光標記的siRNA對照)。

  • 質粒轉染 

Plasmid DNA Transfection
 

  • 質粒制備必須去內毒素,不可用TE等buffer溶解,否則會嚴重影響轉染效率。
  • Namipo轉染試劑使用前請用振蕩器輕微震蕩5s混勻,如有少量沉淀屬于正常現象,產品呈透明或乳白色均不影響產品品質。
  • 根據使用量(參考附表三)將Namipo轉染試劑(μl)與無內毒素水溶質粒(μg) 1:1混合,移液器輕輕吹打5-10次充分混勻,室溫或4℃孵育10分鐘。
  • 按照使用量將Namipo-Plasmid DNA復合物加入培養基,輕輕吹打或8字搖勻。培養基無需特殊處理,24小時內建議不要換液。
  • 轉染后檢測,若所轉染的質粒帶有熒光蛋白,可在轉染后24-48小時后觀察熒光,觀察轉染效率。若質粒不帶熒光,轉染后24-48小時可通過qPCR或wb等方法檢測轉染基因的表達水平,(NamipoTM  轉染試劑試劑盒會贈送10ug 無內毒素的copGFP熒光蛋白質粒)。

 
 
備注:
附表一 配置20μM溶液用量表


摩爾量 nmol 0.5 1 2 5 10 20
溶解量(μL) 25 50 100 250 500 1000
備注;siRNA合成后一般為干粉,根據提供的nmol數參照表格配置,過程中需要 RNase-Free環境。
附表二 各種濃度siRNA用量表
孔板 100nM 50nM 30nM 20nM 10nM 每孔中總體積
96-well 0.5μL 0.25μL 0.15μL 0.1μL 0.05μL 100μL
24-well 2.5μL 1.25μL 0.75μL 0.5μL 0.25μL 500μL
12-well 5μL 2.5μL 1.5μL 1μL 0.5μL 1000μL
6-well 10μL 5μL 3μL 2μL 1μL 2000μL
 

備注;體積為20μM母液使用量
附表三 轉染體積參考表


培養規格
皿/板
表面積
(cm2)
培養液
(μL)
D N A
(μg)
Namipo
(μL)
R N A i
(μL)
Namipo
(μL)
種板密度*(個/孔)
96 well 0.3 100μL 0.3μg 0.3μL 0.3μL 0.3μL 0.5-4 *104
24 well 2 500μL 1.5μg 1.5μL 1.5μL 1.5μL 0.5-2*105
12 well 4 1ml 3.0μg 3.0μL 3.0μL 3.0μL 1-3*105
6  well 10 2ml 6.0μg 6.0μL 6.0μL 6.0μL 2-3*105
60  mm 20 4ml 12.0μg 12.0μL 12.0μL 12.0μL 8-10*105
備注;可根據細胞轉染難易程度上下調整0.3倍的使用量。推薦的siRNA 轉染濃度為60nM,即表格中數值。


效果反饋

\  
Nanop si  VB2細胞 中科院健康所  
\  
Nanop Raw264.7 siRNA 交大醫學院  
\  
Nanop si A549      華西客戶  


文獻參考


 

1.LncRNA-UCA1 exerts oncogenic functions in non-small cell lung cancer by targeting miR-193a-3p

W Nie, H Ge, X Yang, X Sun, H Huang, X Tao, W Chen… - Cancer letters, 2016 – Elsevier

2.Effects of BMAL1–SIRT1-positive cycle on estrogen synthesis in human ovarian granulosa cells: an implicative role of BMAL1 in PCOS

J Zhang, J Liu, K Zhu, Y Hong, Y Sun, X Zhao, Y Du… - Endocrine, 2016 - Springer

3. Upregulation of the Non-Coding RNA OTUB1-isoform 2 Contributes to Gastric Cancer Cell Proliferation and Invasion and Predicts Poor Gastric Cancer …

Y Wang, Q Zhang, W Weng, Y Wu, Y Yang… - … journal of biological …, 2016 - ncbi.nlm.nih.gov

4. OTUB1 promotes tumor invasion and predicts a poor prognosis in gastric adenocarcinoma

W Weng, Q Zhang, M Xu, Y Wu, M Zhang, C Shen… - Am J Transl Res, 2016 - ajtr.org

5.TGR5 activation suppressed S1P/S1P2 signaling and resisted high glucose-induced fibrosis in glomerular mesangial cells

Z Yang, F Xiong, Y Wang, W Gong, J Huang… - Pharmacological …, 2016 - Elsevier

6.Long non-coding RNA LOC389641 promotes progression of pancreatic ductal adenocarcinoma and increases cell invasion by regulating E-cadherin in a …

S Zheng, H Chen, Y Wang, W Gao, Z Fu, Q Zhou… - Cancer letters, 2016 - Elsevier

7.Cyclin Y regulates the proliferation, migration, and invasion of ovarian cancer cells via Wnt signaling pathway

H Liu, H Shi, Q Fan, X Sun - Tumor Biology, 2016 - Springer

8.Dynamic quantitative proteomics characterization of TNF-α-induced necroptosis

Y Wang, ZH Huang, YJ Li, GW He, RY Yu, J Yang… - Apoptosis, 2016 - Springer

9.Inhibition of cyclooxygenase-2 alleviates liver cirrhosis via improvement of the dysfunctional gut-liver axis in rats

JH Gao, SL Wen, H Tong, CH Wang… - American Journal of …, 2016 - Am Physiological Soc
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11. CXCL12/CXCR4 Axis Regulates Aggrecanase Activation and Cartilage Degradation in a Post-Traumatic Osteoarthritis Rat Model

W Lu, J Shi, J Zhang, Z Lv, F Guo, H Huang… - International Journal of …, 2016 - mdpi.com
……
 
 
 
 


 


 


 


 

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