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PAXgene Tissue STABILIZER

For stabilization of tissue specimens previously fixed in PAXgene Tissue FIX
  • Preservation of both morphology and biomolecules
  • Tissue can be stored and archived for later processing
  • RNA, miRNA, DNA and/or proteins from one sample
  • Can be used to fill a tissue processor at position 1
The PAXgene Tissue STABILIZER Concentrate is intended for the stabilization and storage of tissue specimens. This stabilizing agent is part of the PAXgene Tissue System and must be used in conjunction with the PAXgene Tissue FIX Container. The PAXgene Tissue System additionally includes the PAXgene Tissue RNA/miRNA Kit for the isolation of total RNA, including miRNA, and the PAXgene Tissue DNA Kit for DNA. Supplementary protocols are available for other applications, including purification of proteins.
Cat No./ID: 765512
PAXgene Tissue STABILIZER Concentrate (150 ml)
$258.00
Order Product
8 bottles of PAXgene Tissue Stabilizer concentrate, for 4 liters of PAXgene Tissue STABILIZER
For Research Use Only. Not for use in diagnostics procedures. No claim or representation is intended to provide information for the diagnosis, prevention, or treatment of a disease.
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Preservation of tissue morphology.
Tissue was fixed and stabilized with the PAXgene Tissue FIX Container (50 ml); 4 µm sections of PAXgene Tissue fixed, paraffin-embedded (PFPE) tissue were stained with hematoxylin and eosin (HE). [A] Rat liver, [B] rat kidney, [C] rat intestine. Overview at 40x, details at 400x magnification.
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Yield of high-quality, high molecular weight DNA from 12 different PAXgene Tissue-fixed (PF) tissue types, processed on the QIAcube.

Twelve different PAXgene Tissue-fixed (PF) rat tissue types, including fibrous or fatty, DNA-rich and DNA-poor tissues, processed with the PAXgene Tissue DNA Kit on the QIAcube in one single run. DNA yield per 10 mg tissue.

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Absorbance ratio of high-quality, high molecular weight DNA from 12 different PAXgene Tissue-fixed, paraffin-embedded (PF) tissue types, processed on the QIAcube.

Twelve different PAXgene Tissue-fixed (PF) rat tissue types, including fibrous or fatty, DNA-rich and DNA-poor tissue, processed with the PAXgene Tissue DNA Kit on the QIAcube in a single run. Ratio of absorbance at 260 and 280 nm.

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Comparison of manual and automated procedure: DNA yield from sections of PAXgene Tissue-fixed, paraffin-embedded (PFPE) tissue.

DNA yield from 3 sections (thickness 10 µm) each of 8 different PAXgene Tissue-fixed, paraffin-embedded (PFPE) rat tissue types processed in triplicate with the PAXgene Tissue DNA Kit on the QIAcube or manually.

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Comparison of manual and automated procedure: Absorbance ratio from sections of PAXgene Tissue-fixed, paraffin-embedded (PFPE) tissue.

Ratio of absorbance at 260 and 280 nm from 3 sections (thickness 10 µm) each of 8 different PAXgene Tissue-fixed, paraffin-embedded (PFPE) rat tissue types processed in triplicate with the PAXgene Tissue DNA Kit on the QIAcube or manually.

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DNA without chemical modification can be used for demanding downstream applications.

Multiplex and long-range PCR of DNA from PAXgene Tissue-fixed, paraffin-embedded (PFPE) human colorectal cancer tissue (modified according to Viertler et al., (2012) A new technology for stabilization of biomolecules in tissues for combined histological and molecular analyses. J. MOl. Diagn. 14, 458). [A] Multiplex PCR of 8 different genomic DNA fragments ranging from 22 to 955 bp. [B] Long-range PCR of a 5 kb genomic DNA fragment.

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Protein extracted from PFPE tissue is suitable for two-dimentional gel electrophoresis.

Non-malignant human duodenum tissue was divided into 3 samples and either flash-frozen in liquid nitrogen (cryo), or prepared as PFPE or FFPE tissue. Proteins from cryo and PFPE tissue were extracted in 2D buffer (30 mM Tris-HCl, pH 8.8, 7 M urea, 2 M thiourea, 4% CHAPS, 75 mM DTT) supplemented with protease inhibitor. Proteins from FFPE tissue were extracted in EXB Plus buffer supplemented with protease inhibitor, precipitated with acetone and resuspended in 2D buffer (as described in Guendisch et al. 2013). Samples (150 µg) were separated by 2-D PAGE. Data kindly provided by Karl-Friedrich Becker, Technical University of Munich, Germany.

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Agarose gel electrophoresis of high-quality, high molecular weight DNA from 12 different PAXgene Tissue-fixed (PF) tissue types, processed on the QIAcube.

Twelve different PAXgene Tissue-fixed (PF) rat tissue types, including fibrous or fatty, DNA-rich and DNA-poor tissue, processed with the PAXgene Tissue DNA Kit on the QIAcube in one single run. DNA from each tissue type (300 ng) on agarose gel electrophoresis using 1% TBE buffered gels; M: markers.

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Comparison of manual and automated procedure: Agarose gel electrophoresis from sections of PAXgene Tissue-fixed, paraffin-embedded (PFPE) tissue.

Agarose gel electrophoresis from 3 sections (thickness 10 µm) each of 8 different PAXgene Tissue-fixed, paraffin-embedded (PFPE) rat tissue types processed in triplicate with the PAXgene Tissue DNA Kit on the QIAcube and manually. DNA from each replicate and tissue type (200 ng) on agarose gel electrophoresis using 1% TBE buffered gels; M: markers.

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Detection of nondegraded, immunoreactive phosphoproteins from human clinical PFPE tissue.

Non-malignant human uterus, breast, prostate and bladder tissue specimens were divided into 3 samples and either flash-frozen in liquid nitrogen (cryo), PAXgene Tissue-fixed, paraffin-embedded (PFPE) or formalin-fixed, paraffin-embedded (FFPE). Proteins from cryo, PFPE and FFPE tissues were extracted using the extraction buffer EXB Plus (Qproteome FFPE Tissue Kit; described in Ergin et al. 2010, Guendisch et al. 2013 and PAXgene Tissue supplementary protocols). SDS-PAGE and Western blot analysis were performed with 15 µg protein and the indicated antibodies. Data kindly provided by Karl-Friedrich Becker, Technical University of Munich, Germany.

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High concordance of miRNA expression between total RNA isolated from PFPE and fresh-frozen tissue.

RNA, including miRNA, was purified from mirrored human breast cancer tissue fresh frozen in liquid nitrogen using the QIAGEN miRNeasy Kit, or from sections of PAXgene Tissue-fixed, paraffin-embedded (PFPE) tissue using the PAXgene Tissue RNA/miRNA Kit. Shown is a scatterplot of CT values from different single miRNA-specific RT-qPCR assays using the QIAGEN miScript System: miR9, -10a, -10b, -29a, -103, -125b, -143, -145, -192 and miScript PCR controls RNUA1, RNU5A, RNU6B, SNORD25, SCARNA19, SNORA73A; R2: coefficient of determination.

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RNA purified without chemical modification from PFPE tissue using the PAXgene Tissue RNA/miRNA Kit.

SYBR Green real-time RT-qPCR was performed with 10 ng RNA from cryopreserved, formalin-fixed, paraffin-embedded (PPFE) or PAXgene Tissue-fixed, paraffin-embedded (PFPE) rat tissue (modified according to Groelz et al. 2013). Depicted are the average delta-CT values (delta-CT = CT[FFPE] – CT[cryo] or delta-CT = CT[PFPE] – CT[cryo]) from 6 different assays with amplicons ranging from 109 to 465 bp.

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Immunohistochemistry (IHC) staining with the PAXgene Tissue System gives comparable results to formalin-fixed tissue.

Human palatine tonsil tissue was fixed in PAXgene Tissue FIX or with neutral-buffered formalin and embedded in paraffin. Primary antibodies to the indicated antigens were linked to a streptavidin-peroxidase conjugate by a biotinylated secondary antibody (LSAB method). Sectiosn were counterstained with hematoxylin. PFPE: PAXgene Tissue-fixed, paraffin-embedded. FFPE: formalin-fixed, paraffin-embedded.

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H&E staining with the PAXgene Tissue System gives results comparable to formalin-fixed tissue.

Human tissue samples were divided into 2 sub-samples. One sub-sample was fixed with PAXgene Tissue FIX and the other was fixed with neutral-buffered formalin. The fixed tissues were embedded in paraffin, sectioned and stained with hematoxylin and eosin. PFPE: PAXgene Tissue-fixed, paraffin-embedded. FFPE: formalin-fixed, paraffin-embedded.

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The PAXgene Tissue FIX Container (50 ml) and PAXgene Tissue STABILIZER workflow.

Single-chamber container for fixation and stabilization of a single, larger or multiple, smaller tissue samples.

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High-quality DNA from PFPE tissue with preserved morphology.

[A] Hematoxylin and eosin (H&E) staining of PAXgene Tissue-fixed, paraffin-embedded (PFPE) human colorectal cancer tissue and [B] DNA on agarose gel electrophoresis using 0.8% TBE buffered gels with 200 ng genomic DNA isolated in triplicate from 5 cases (#1–5) of human PFPE colorectral cancer. M: markers.

Performance
The PAXgene Tissue STABILIZER Concentrate serves to stabilize and preserve tissue morphology and the integrity of biomolecules without destructive crosslinking and degradation found in formalin-fixed tissues (see H&E staining with the PAXgene Tissue System gives results comparable to formalin-fixed tissue and Immunohistochemistry (IHC) staining with the PAXgene Tissue System gives comparable results to formalin-fixed tissue). While the stabilizing action of this agent is designed for tissues that have been previously fixed in PAXgene Tissue FIX, appropriately diluted PAXgene Tissue STABILIZER Concentrate can be used in paraffin-embedding with a tissue processor.

Enhanced storage
When tissues previously fixed in PAXgene Tissue FIX are stored in PAXgene Tissue STABILIZER, nucleic acids, proteins and morphology are stable for up to 7 days at room temperature (15–25°C) or 4 weeks at 2–8°C. Tissue samples can be stored in PAXgene Tissue STABILIZER for longer periods at –20°C (–15°C to –30°C) or –80°C (–65°C to –90°C) without negative effects on the morphology of the tissue or the integrity of analytes.

Note: Specifications for storage conditions in PAXgene Tissue STABILIZER were determined using animal tissues. Storage at 2–8°C for more than 4 weeks must be validated for each tissue type.

Note: For storage at –20°C (–15°C to –30°C) or –80°C (–65°C to –90°C), use cryogenic vials with screw caps filled with PAXgene Tissue STABILIZER. For safety reasons, note that PAXgene Tissue STABILIZER contains 70% ethanol by volume.


High-quality biomolecule stabilization

Nucleic acids and proteins of tissues stored in PAXgene Tissue STABILIZER or as PAXgene Tissue-fixed, paraffin-embedded (PFPE) samples are stabilized and can be purified using the corresponding PAXgene Tissue Kit for RNA and miRNA or DNA. Use the Qproteome FFPE Tissue Kit for purification of proteins. Purified biomolecules are of high quality (see High-quality DNA from PFPE tissue with preserved morphology) and unmodified (see DNA without chemical modifications can be used for demanding downstream applications). The purified biomolecules are highly suited for a range of demanding downstream applications (see High concordance of miRNA expression between total RNA isolated from PFPE and fresh-frozen tissueDetection of nondegraded, immunoreactive phosphoproteins from human PFPE tissue and RNA purified without chemical modification from PFPE tissue using the PAXgene Tissue RNA/miRNA Kit).
Principle
The formalin-free PAXgene fixation and stabilization products can be used as an alternative to traditional histology methods for tissue fixation and analysis because they enable comparable processing of tissue samples without the crosslinking and degradation observed in formalin-based systems. The PAXgene Tissue STABILIZER enables stabilization of tissue specimens previously fixed in PAXgene Tissue FIX Container. The system facilitates preservation of tissue morphology and stabilization of biomolecules so that histological and molecular analyses, including RT-PCR, qPCR and next-generation sequencing, can be performed on the same tissue specimen.

Procedure
PAXgene Tissue STABILIZER Concentrate is diluted with ethanol to make PAXgene Tissue STABILIZER Reagent, which is designed to be used with tissues that have been fixed in PAXgene Tissue FIX. After fixation of tissue samples with the PAXgene Tissue FIX Containers for 2–72 hours, the fixative in the container is removed and replaced by PAXgene Tissue STABILIZER. Stabilized samples can be stored long-term in the STABILIZER or embedded in paraffin for histological studies. Nucleic acids and proteins can be isolated from the stabilized samples before or after embedding in paraffin. PAXgene Tissue STABILIZER Reagent can also be used in paraffin embedding with a tissue processor (see The PAXgene Tissue FIX Container (50 ml) and PAXgene Tissue STABILIZER workflow).
Applications
PAXgene Tissue-fixed (PF) and PAXgene Tissue-fixed, paraffin-embedded (PFPE) tissue samples can be used for:
  • Histopathological staining, including hematoxylin & eosin (H&E), periodic acid schiff (PAS), resorcin fuchsin, sirius red and Gomori
  • Immunhistochemical staining
  • in situ hybridization

 

Nucleic acids purified from PF and PFPE tissue samples can be used for demanding downstream applications, including:
  • RNA and miRNA profiling
  • Long-range and multiplex PCR
  • Next-generation sequencing

 

Proteins purified from PF and PFPE tissue samples can be used in a range of downstream applications, including:
  • Western blotting
  • Reverse-phase protein microarrays
  • MALDI imaging mass spectrometry
  • 2-D gel electrophoresis


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Brochures & Guides (6)
For isolation and purification of genomic DNA from tissue samples stabilized using the PAXgene Tissue System 

 


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For isolation and purification of total RNA, including miRNA, from tissue samples fixed and stabilized using the PAXgene Tissue System

 


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For extraction of full-length proteins from PFPE tissue using the Qproteome FFPE Tissue Kit

 


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For isolation and purification of intracellular RNA from tissue samples stabilized using the PAXgene Tissue System

 


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Moving toward excellence and standardization in tissue collection and fixation

 


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FAQs (34)
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Scientific Posters (15)
Long et al., ADAPT 2012

 


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Hesse et al., AACR-NCI-EORTC 2011

 


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Groelz et al., AACR 2010

 


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Groelz et al., AMP 2010 

 


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Groelz et al., AMP 2008 

 


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Groelz et al., AMP 2009 

 


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Groelz et al., ISBER 2012

 


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Groelz et al., BRN Symposium 2012 

 


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Groelz et al., ECP 2012

 


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Groelz et al., AMP 2009

 


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Groelz et al., 3rd Annual Oncology Biomarkers 2011

 


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Groelz et al., GTEx Meeting 2014

 


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Groelz et al., AMP 2014

 


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Groelz et al., BRN Symposium 2011 

 


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Groelz et al., ECP 2014

 


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Kit Handbooks (2)
For collection, fixation, and stabilization of multiple small tissue samples or a single large tissue sample

 


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For stabilization of tissue specimens fixed in PAXgene Tissue FIX Container

 


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Supplementary Protocols (13)

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References
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