迅速稳定组织中的DNA、RNA和蛋白质

  • 无需液氮或干冰
  • 立即方便地稳定收集到的组织
  • 可长期储存组织,用于后续分析
  • 标准的样本制备流程,用于系统生物学研究

Allprotect Tissue Reagent可在室温下立即稳定组织样本中的DNA、RNA和蛋白质。这样可以稳定DNA、RNA和蛋白质,以保证可靠的下游分析。稳定后的组织可在15–25°C运输7天,2–8°C储存长达12个月,在–20°C或–80°C可储存更长时间。

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Allprotect Tissue Reagent (100 ml)
100 ml Allprotect Tissue Reagent, Allprotect Reagent Pump
76405
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The Allprotect Tissue Reagent is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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Preservation of protein activity.|Prevention of gene induction.|Prevention of RNA degradation.|Intact protein for western blotting.|Prevention of protein degradation.|Mechanism of Allprotect stabilization.|
Rat liver was stored in Allprotect Reagent before total protein was purified using the Qproteome Mammalian Protein Prep Kit. Acidic phosphatase activity was similar to that of protein from control liver stored in liquid nitrogen. Each bar shows the mean from two liver pieces assayed in duplicate.|Rat lung was stored at 25°C in Allprotect Reagent or in PBS for the indicated times prior to real-time RT-PCR analysis of c-fos expression. Transcript levels relative to those of liquid nitrogen stabilized lung were calculated. The data show that Allprotect Reagent prevents c-fos induction.|Rat intestine was stored at 25°C in Allprotect Reagent or in PBS for the indicated times prior to real-time RT-PCR analysis of Madh7 expression. Transcript levels relative to those of liquid nitrogen stabilized intestine were calculated. The data show that Allprotect Reagent prevents RNA degradation.|Various rat tissues were stored in Allprotect Reagent at 25°C for 5 days prior to western blot analysis of ERK2. Blot intensities were similar to those from liquid nitrogen stabilized tissues, indicating no protein degradation.|Human intestinal mucosa samples were stored for 1 day in Allprotect Reagent (4°C) or in liquid nitrogen, and then analyzed by mass spectrometry. The Allprotect samples gave similar peak patterns for [A] <10 kDa and [B] 10–20 kDa protein as the liquid nitrogen samples, indicating stabilization of the protein content. Example analyses are shown. (Data kindly provided by Indivumed GmbH, Hamburg, Germany).|Each Allprotector molecule possesses contact points that allow multiple Allprotector molecules to envelop and protect DNA, RNA, and protein. Upon tissue lysis, Allprotector molecules become diluted and fall apart, releasing DNA, RNA, and protein.|
Performance
使用Allprotect Tissue Reagent即刻稳定采集的组织可保留DNA、RNA和蛋白质在体内的性质,保证了可靠的下游分析,稳定的组织可在室温(15–25°C)下储存长达7天(参见"Intact protein for western blotting"、"Prevention of protein degradation"和"Preservation of protein activity")。稳定RNA十分重要,因为转录水平会在组织采集后快速变化(参见"Prevention of gene induction"和"Prevention of RNA degradation")。使用Agilent 2100 Bioanalyzer分析Allprotect Tissue Reagent稳定组织得到的RNA,RIN值较高,表明RNA较完整。Allprotect Tissue Reagent还可稳定microRNA、甲基化的基因组DNA和磷酸化蛋白。
Principle

将刚采集的组织浸没在Allprotect Tissue Reagent中,可即刻稳定DNA、RNA和蛋白质,并保存其在体内的特性。Allprotect Tissue Reagent稳定的组织样本可长期储存,无需冷冻,在室温下可储存长达7天,或在2–8°C储存长达12个月。

该试剂是QIAGEN系统生物学样本制备技术产品线的一部分,可与AllPrep DNA/RNA/Protein Mini Kit配合使用,从同一个珍贵的样本中同时纯化DNA、RNA和蛋白质。Allprotect稳定的组织可用QIAGEN的DNeasy、QIAamp、RNeasy和Qproteome等试剂盒纯化DNA、RNA或蛋白质。用于酶分析的功能性蛋白可使用Qproteome Mammalian Protein Prep Kit从Allprotect稳定的组织中纯化获得。无需在纯化前将样本分为3份,因此可最大程度的回收DNA、RNA和蛋白质。此外,纯化过程无需加入苯酚和丙酮等有机溶剂。DNA、RNA和蛋白质皆来自同一个样本,减少了从不同样本中制备这些待分析物可能带来的差异。

QIAGEN还提供分别纯化DNA、RNA和蛋白质的产品(见表)。包括Qproteome Mammalian Protein Prep Kit,可从Allprotect稳定的组织中纯化功能蛋白。使用Qproteome Mammalian Protein Prep Kit从Allprotect稳定的组织中纯化的蛋白质可用于多种下游应用。

可与Allprotect Tissue Reagent配合使用的QIAGEN试剂盒
待纯化的分析物推荐的试剂盒
基因组DNA、总RNA和总蛋白质 AllPrep DNA/RNA/Protein Mini Kit
基因组DNA和总RNA AllPrep DNA/RNA Micro Kit
AllPrep DNA/RNA Mini Kit
基因组DNA DNeasy Blood & Tissue Kit
QIAamp DNA Mini Kit
总RNA RNeasy Plus Micro Kit*(适用于小的组织样本)RNeasy Mini Kit(适用于易裂解组织)
RNeasy Plus Mini Kit (适用于易裂解组织)
RNeasy Fibrous Tissue Mini Kit* (适用于富含纤维的组织)
RNeasy Lipid Tissue Mini Kit(适用于富含脂肪的组织)
RNeasy Microarray Tissue Mini Kit(适用于微芯片分析中的组织样本)
总蛋白质 Qproteome Mammalian Protein Prep Kit
* 试剂盒中含有gDNA Eliminator柱。
Procedure

将刚采集的组织浸没在Allprotect Tissue Reagent中,以即刻稳定DNA、RNA和蛋白质(参见"Mechanism of Allprotect stabilization")。稳定过程在室温下即可进行,无需使用危险的液氮或干冰。使用Allprotect Tissue Reagent即刻稳定采集的组织可保留DNA、RNA和蛋白质在体内的性质,保证了可靠的下游分析。

Allprotect Tissue Reagent稳定的组织可在室温下储存长达7天,在2–8°C的条件下储存长达12个月, DNA、RNA和蛋白质不会降解。在–20°C或–80°C可储存更长时间。

Applications

使用AllPrep DNA/RNA/Protein Mini Kit可从Allprotect稳定的组织中同时纯化DNA、RNA和蛋白质。将Allprotect和AllPrep样本制备技术配合,可标准化系统生物学研究中的样本制备流程。使用AllPrep DNA/RNA/Protein Mini Kit纯化得到的基因组DNA和总RNA在real-time PCR和RT-PCR等下游应用中表现出色。使用该试剂盒纯化得到的总蛋白质适用于SDS-PAGE和Western印迹。

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Kit Handbooks
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For immediate stabilization of DNA, RNA, and proteins in animal and human tissues, and subsequent storage
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