Allprotect Tissue Reagent
For immediate stabilization of DNA, RNA, and protein in tissues
- No need for liquid nitrogen or dry ice
- Immediate, convenient stabilization of harvested tissues
- Long-term storage of tissues for later analysis
- Standardized sample preparation for systems biology
Allprotect Tissue Reagent provides immediate stabilization of DNA, RNA, and protein in tissue samples at room temperature. This preserves the in vivo profile of DNA, RNA, and proteins, allowing reliable downstream analysis. Stabilized tissues can be transported at 15–25°C for up to 7 days, or stored at 2–8°C for up to 12 months. For longer storage, stabilized tissues can be archived at –20°C or –80°C.
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Allprotect Tissue Reagent (100 ml)
100 ml Allprotect Tissue Reagent, Allprotect Reagent Pump
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76405
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Allprotect Tissue Reagent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。
保持蛋白活性。|防止基因诱导。|防止RNA降解。|完整的蛋白可用于Western印迹。|防止蛋白质降解。|Allprotect稳定的机理。|
大鼠肝脏储存在Allprotect Reagent中,然后使用Qproteome Mammalian Protein Prep Kit纯化总蛋白。酸性磷酸酶活性与储存于液氮中的肝脏(对照)相似。图中的柱为平行分析两种保存方法的肝脏组织的蛋白活性平均数值。|将大鼠肺储存在Allprotect Reagent或PBS中,按图中所示时间存放于25°C环境中。然后进行real-time RT-PCR,分析c-fos的基因表达。分别计算用液氮稳定的肺样本的转录水平。数据表明Allprotect Reagent能够防止c-fos基因诱导。|将大鼠的小肠储存在Allprotect Reagent或PBS中,按图中所示时间存放于25°C环境中。然后进行real-time RT-PCR,分析Madh7的基因表达。分别计算用液氮稳定的小肠样本的转录水平。数据表明Allprotect Reagent能够防止RNA降解。|将大鼠的多种组织储存在Allprotect Reagent中,在25°C环境中储存5天。然后对ERK2进行Western印迹分析。其印迹强度与液氮稳定的组织的印迹强度相当,表明没有出现蛋白质降解。|将人体肠道粘膜样品在Allprotect Reagent (4°C)或液氮中储存1天,然后进行质谱分析。对于[A] <10 kDa的蛋白和[B] 10–20 kDa的蛋白,Allprotect中样本的图形与液氮中样本图形类似,表明蛋白质已被稳定。图中数据为德国汉堡Indivumed GmbH提供。|每个Allprotector分子具有多个允许Allprotector分子包被并保护DNA、RNA和蛋白质的接触点。组织裂解时,Allprotector分子被稀释、分解,释放出DNA、RNA和蛋白质。|
性能
Immediate stabilization of harvested tissues in Allprotect Tissue Reagent preserves the in vivo profile of DNA, RNA, and proteins. This enables reliable downstream analysis even with tissues stored at room temperature (15–25°C) for up to 7 days (see figures " Intact protein for western blotting", " Prevention of protein degradation", and " Preservation of protein activity"), Stabilization is especially critical for RNA, since transcript levels change rapidly after tissue harvesting (see figures " Prevention of gene induction" and " Prevention of RNA degradation"). Agilent 2100 Bioanalyzer analysis of RNA from Allprotect stabilized tissues gives high RIN values, indicating intact RNA. Allprotect Tissue Reagent is also able to stabilize molecules of increasing interest such as miRNA, methylated genomic DNA, and phosphorylated protein.
原理
Freshly harvested tissue is submerged in Allprotect Tissue Reagent to immediately preserve the in vivo profile of DNA, RNA, and protein. Tissue samples in Allprotect Tissue Reagent are stabilized for long-term storage without freezing and can be stored at room temperature for up to 7 days or at 2–8°C for up to 12 months.
The reagent is part of QIAGEN’s range of sample technologies for systems biology, and can be used in combination with the AllPrep DNA/RNA/Protein Mini Kit, which allows simultaneous purification of DNA, RNA, and protein from the same precious sample. Allprotect stabilized tissues are also compatible with QIAGEN kits for purification of DNA, RNA, or protein, such as DNeasy, QIAamp, RNeasy, and Qproteome Kits. Functional protein for enzyme assays can be purified from Allprotect stabilized tissues using the Qproteome Mammalian Protein Prep Kit. There is no need to split the sample into 3 prior to purification, allowing maximum recovery of DNA, RNA, and protein. In addition, no organic solvents such as phenol and acetone are required. DNA, RNA, and protein are all prepared from the same source, eliminating the variation inherent in preparing these analytes from different samples.
QIAGEN sample technologies for individual purification of DNA, RNA, and protein are also available (see table). These include the Qproteome Mammalian Protein Prep Kit, which allows purification of functional protein from Allprotect stabilized tissues. Protein purified from Allprotect stabilized tissue with the Qproteome Mammalian Protein Prep Kit is suitable for all downstream applications.
| Genomic DNA, total RNA, and total protein |
AllPrep DNA/RNA/Protein Mini Kit |
| Genomic DNA and total RNA |
AllPrep DNA/RNA Micro Kit
AllPrep DNA/RNA Mini Kit
AllPrep DNA/RNA/miRNA Universal Kit |
| Genomic DNA |
DNeasy Blood & Tissue Kit
QIAamp DNA Mini Kit |
| Total RNA |
RNeasy Plus Micro Kit* (for small tissue samples)
RNeasy Mini Kit (for easy-to-lyse tissues)
RNeasy Plus Mini Kit* (for easy-to-lyse tissues)
RNeasy Fibrous Tissue Mini Kit (for fibrous tissues)
RNeasy Lipid Tissue Mini Kit (for fatty tissues)
RNeasy Microarray Tissue Mini Kit (for tissue samples for microarray analysis) |
| Total protein |
Qproteome Mammalian Protein Prep Kit |
操作流程
Freshly harvested tissue is submerged in Allprotect Tissue Reagent to immediately stabilize DNA, RNA, and protein (see figure "Mechanism of Allprotect stabilization"). Stabilization takes place at room temperature, eliminating the use of hazardous liquid nitrogen or dry ice. Immediate stabilization of harvested tissues in Allprotect Tissue Reagent preserves the in vivo profile of DNA, RNA, and proteins, enabling reliable downstream analysis.
Tissues stabilized in Allprotect Tissue Reagent can be stored at room temperature for up to 7 days or at 2–8°C for up to 12 months without showing any degradation of DNA, RNA, and protein. For longer storage, stabilized tissues can be archived at –20°C or –80°C.
应用
DNA, RNA, and protein can be simultaneously purified from Allprotect stabilized tissues using the AllPrep DNA/RNA/Protein Mini Kit. The combination of Allprotect and AllPrep sample technologies standardizes sample preparation for systems biology. Genomic DNA and total RNA purified using the AllPrep DNA/RNA/Protein Mini Kit deliver optimal results in all downstream applications. These include real-time PCR and RT-PCR analyses. Protein purified from Allprotect stabilized tissue is denatured and suitable for SDS-PAGE and western blotting.
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FAQ ID -1571
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FAQ ID -1572
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FAQ ID -1573
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FAQ ID -1574
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FAQ ID -1575
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For immediate stabilization of DNA, RNA, and proteins in animal and human tissues, and subsequent storage
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图片
保持蛋白活性。
大鼠肝脏储存在Allprotect Reagent中,然后使用Qproteome Mammalian Protein Prep Kit纯化总蛋白。酸性磷酸酶活性与储存于液氮中的肝脏(对照)相似。图中的柱为平行分析两种保存方法的肝脏组织的蛋白活性平均数值。
防止基因诱导。
将大鼠肺储存在Allprotect Reagent或PBS中,按图中所示时间存放于25°C环境中。然后进行real-time RT-PCR,分析c-fos的基因表达。分别计算用液氮稳定的肺样本的转录水平。数据表明Allprotect Reagent能够防止c-fos基因诱导。
防止RNA降解。
将大鼠的小肠储存在Allprotect Reagent或PBS中,按图中所示时间存放于25°C环境中。然后进行real-time RT-PCR,分析Madh7的基因表达。分别计算用液氮稳定的小肠样本的转录水平。数据表明Allprotect Reagent能够防止RNA降解。
完整的蛋白可用于Western印迹。
将大鼠的多种组织储存在Allprotect Reagent中,在25°C环境中储存5天。然后对ERK2进行Western印迹分析。其印迹强度与液氮稳定的组织的印迹强度相当,表明没有出现蛋白质降解。
防止蛋白质降解。
将人体肠道粘膜样品在Allprotect Reagent (4°C)或液氮中储存1天,然后进行质谱分析。对于[A] <10 kDa的蛋白和[B] 10–20 kDa的蛋白,Allprotect中样本的图形与液氮中样本图形类似,表明蛋白质已被稳定。图中数据为德国汉堡Indivumed GmbH提供。
Allprotect稳定的机理。
每个Allprotector分子具有多个允许Allprotector分子包被并保护DNA、RNA和蛋白质的接触点。组织裂解时,Allprotector分子被稀释、分解,释放出DNA、RNA和蛋白质。
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