Allprotect Tissue Reagent

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.

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 andcan be stored at room temperature for up to 7 days or at 2–8°C for up to12 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.

QIAGEN kits compatible with Allprotect Tissue Reagent

* Kit includes gDNA Eliminator columns

Analytes purified	Recommended kit
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

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.

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.