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AllPrep RNA/Protein Kit

从细胞中同时纯化RNA和蛋白质

  • 从同一样本中纯化获得高品质RNA和蛋白质
  • 采用离心柱法快速纯化
  • 采用较少的细胞高效的进行样本制备
  • 获得高品质的RNA和天然蛋白

AllPrep RNA/Protein Kit可以从同一个细胞培养样本中同时纯化总RNA和天然蛋白。

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Cat No./ID: 80404
AllPrep RNA/Protein Kit (50)
50 AllPrep Mini Spin Columns, 50 RNeasy Mini Spin Columns, 50 Protein Cleanup Mini Spin Columns, Collection Tubes, RNase-Free Reagents and Buffers

AllPrep RNA/Protein Kit适用于分子生物学应用。该产品不适用于疾病的诊断、预防或治疗。


Product Details

9
Sensitivity in real-time RT-PCR.
RNA was purified from HeLa cells using the AllPrep RNA/Protein Kit and reverse-transcribed. Serial dilutions of cDNA equivalent to 100 pg to 100 ng of total RNA were analyzed using a real-time RT-PCR assay for human GSK3B (glycogen synthase kinase 3 beta). Reactions were performed in duplicate. RNA purified using the RNeasy Mini Kit showed similar CT values and reproducibility (data not shown). NTC: No template control.
9
AllPrep RNA/Protein procedure.
 
9
Purification of functional protein.
HeLa cells in a 96-well plate were transfected with a β-galactosidase-reporter plasmid. After 48 hours, protein was purified and β-galactosidase activity was assayed (yellow indicates enzyme activity). Protein was purified using either the AllPrep RNA/Protein Kit (AllPrep) or the RNeasy Mini Kit (cell lysate was passed through an RNeasy spin column, and protein was precipitated from the flow-through) (Other). Each column of wells shows triplicate results from one cell sample.
9
Highly intact RNA and high-quality protein from the same cell sample.

Total RNA was purified from HeLa cells and analyzed on the Agilent 2100 Bioanalyzer. The rRNA ratio (28S/18S) was 2.04. Total protein was purified from HeLa cells and analyzed by western blot. Analysis of duplicate protein samples with antibodies against Lamin (numclear protein) and TIM23 (mitochondrial protein) demonstrates that the total protein fraction contains proteins from subcellular compartments.

Performance
AllPrep RNA/Protein Kit适用于从同一个细胞培养样本中同时纯化总RNA和天然蛋白。使用Agilent 2100 Bioanalyzer对纯化的RNA进行分析通常获得的RIN值为10,表明RNA十分完整。纯化的天然蛋白包含所有细胞蛋白(参见"Highly intact RNA and high-quality protein from the same cell sample")。AllPrep RNA/Protein Kit纯化得到的RNA适用于real-time RT-PCR(参见"Sensitivity in real-time RT-PCR"),纯化得到的细胞蛋白质可用于酶分析(参见"Purification of functional protein")。
Principle

AllPrep RNA/Protein Kit适用于从同一个细胞培养样本中同时纯化总RNA和天然蛋白。该试剂盒可进行高效的样本制备,使用较少量的细胞即可进行纯化,因此适用于在mRNA和蛋白质水平评估siRNA引起的基因敲除。此外,因为RNA和蛋白质来自同一样本,所以基因敲除分析更加可靠。AllPrep RNA/Protein Kit是QIAGEN RNAi和基因表达分析产品线的一部分,可获得具有高度可重复性且灵敏的结果。该试剂盒包含优化的PCR预混液、即用型试剂、siRNA和转染试剂。该产品使RNAi和基因表达分析工作流程标准化,在生物医学研究的各个领域取得了成功。

Procedure
该试剂盒利用离心柱法,可在40分钟内从同一个细胞样本中纯化RNA和蛋白质(参见"AllPrep RNA/Protein procedure")。AllPrep DNA/RNA/Protein Kit可从培养细胞和组织中同时纯化DNA、RNA和蛋白质(适用于Western印迹,不适用于酶分析)。
Applications

AllPrep RNA/Protein Kit适用于从同一个细胞培养样本中同时纯化总RNA和天然蛋白。纯化得到的RNA和蛋白质在下游应用中表现出色,如real-time RT-PCR、Western印迹和酶分析。(注:待测蛋白的功能需要根据其特性进行检测)。

Specifications

Features
Specifications
应用 PCR, real-time PCR, blotting, microarray
洗脱体积 30–50 µl
规格 Spin column
主要样本类型 Cultured cells
处理 Manual (centrifugation)
纯化总RNA、miRNA、poly A+ mRNA、DNA或蛋白 RNA, native protein
样本量 700 µl
技术 Silica technology
每次运行或制备时间 40 minutes

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