Qproteome FFPE Tissue Kit
For isolation of full-length proteins from formalin-fixed tissues
- Standardized, quantitative extraction from FFPE tissue
- Full-length proteins
- Low sample requirements
Cat. No. / ID: 37623
The Qproteome FFPE Tissue Kit uses innovative proprietary chemistry to reverse the crosslinking process of fomalin fixation and deliver full-length proteins ready for subsequent analysis.
The Qproteome FFPE Tissue Kit delivers efficient protein extraction from FFPE tissue using a standardized protocol for reproducible protein yields (see figure " Standardized and reproducible protein extraction"). Using the Qproteome FFPE Tissue Kit, protein yields are comparable to frozen material, enabling, for example, comparison between different tissues for identification of disease-specific markers (see figure " tissue-specific biomarker identification").
As the standard method for histological tissue preparation, millions of formalin-fixed paraffin-embedded (FFPE) samples are classified and archived each year. Previously, their analysis was limited to histochemical techniques. The Qproteome FFPE Tissue Kit opens this vast, highly valuable resource up to proteomic and protein expression analysis by enabling efficient extraction of full-length proteins suitable for western blotting, protein arrays, and mass spectrometry. For the very first time, proteins or pathway of interest can be investigated in archived FFPE tissues. Probing blots or arrays with multiple antibodies enables simultaneous investigation of proteins, providing a detailed picture of multiple protein expression profiles, for example, in signaling transduction cascades.
Formalin fixation creates chemical crosslinks, stabilizing samples for histological analysis, but rendering any subsequent protein separation (e.g., by SDS-PAGE) impossible. The Qproteome FFPE Tissue Kit uses innovative proprietary chemistry to reverse the crosslinking process and deliver full-length proteins ready for analysis by western blotting or protein array, with subsequent immunodetection (see figure “Compare FFPE Tissue Samples at the Protein Level”).
Sections are deparaffinized and incubated in an Extraction Buffer. After centrifugation, extracted proteins are recovered in the supernatant. Recovered proteins can be mixed directly with sample buffer and loaded onto an SDS-PAGE gel, just like conventional protein samples. No extra expensive equipment or additional processing is required.
The Qproteome FFPE Tissue Kit offers a unique tool for analysis of samples whose pathological status has been determined, but whose precise protein makeup has until now remained inaccessible. Such samples — for example, taken over a course of treatment or from different groups of probands — offer a rich source of information for comparative proteomics and biomarker studies.
Protein extraction enables comparison of histological samples on the protein level. Two breast cancer biopsy samples, both scored 2+ for the receptor tyrosine kinase HER2 by immunohistochemistry, were processed using the Qproteome FFPE Tissue Kit and analyzed by protein lysate array and western blotting. In sample B, HER2 gene amplification was confirmed by FISH (data not shown). The housekeeping gene β-actin was analyzed as a control. Data kindly provided by Karl-Friedrich Becker and Christina Schott, Technical University of Munich, Germany.
|Applications||SDS-PAGE, Western-Blot, mass spectrometry|
|Binding capacity/yield||20–80 µg|
|Start material||Formalin-fixed tissues|
|Sample size||2–3 x 100 mm2 per prep|
|Fractions isolated||One fraction|