Quantitating Protein Expression in Intact Tissue Sections for Cancer Research

8 May 2013

Quantitating the expression of multiple proteins in intact tissue sections provides valuable information about how cancer-related cells function, behave, and respond in context, and how their presence, frequency, and spatial distribution correlate with drug efficacy and clinical outcomes. This information cannot be obtained using methods such as flow cytometry where samples are disaggregated or conventional single-plex IHC where one typically utilizes serial sections. This application note describes several tools that are available for quantitating protein expression in intact tissue sections.

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ProteomicsProteomics is the systemic bioinformatics study of proteins and amino acids, including their structure, size, function and identification. Tools used in proteomics include chromatography, blotting and gels, protein arrays, mass spectrometry and ELISA and associated analysis software. Analyzers and proteomic systems should be sensitive, high resolution, fast and may be automated for high-throughput.Flow Cytometry / Cell CountingFlow cytometers are used to count, sort and examine multiple characteristics of cells. Other cell analysis equipment includes image cytometers, cell counters, fluorescence-activated cell sorters (FACS), magnetic-activated cell sorters (MACS), and a range of flow cytometry assay kits. Flow cytometers can reveal information on cell viability, cell proliferation, apoptosis and cell cycle progression, as well as identify cell populations and intracellular or cell-surface molecules. Additionally, some flow cytometers, known as FACS, have an additional sorting function after analysis. Cell counters and image cytometers count live and dead cell populations and can also conduct cell proliferation assays. Find the best flow cytometers, cell counters and cell sorters in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Protein PurificationProtein purification is a vital step in drug discovery, therapeutics, biotech and life science research. The purification process typically involves subcellular or membrane protein extraction with cell lysis kits, separation of proteins from cell debris by filtration or spin columns, and the isolation of proteins of interest from other proteins and impurities with affinity purification (including fusion protein tags and antibody binding proteins A, G and L), immunoprecipitation or chromatographic methods, such as ion exchange, size exclusion and immobilized metal affinity chromatography. All purification methods come in multiple formats for your laboratory needs, including agarose or magnetic beads, resins, columns and filter plates. Find the best protein purification equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.BiomarkersBiomarkers are biological markers which can be measured and evaluated to indicate a biological state. The use of biomarkers in research and diagnosis can indicate a normal or disease state or drug response of cells / tissues. Biomarkers include genetic markers, cell surface markers such as antigens, antibodies or receptors and secreted molecules such as cytokines. An assay system is required for identification of biomarkers. :Light MicroscopyLight microscopes or optical microscopes are used to visualize microscale objects under magnification, including cells, clinical specimens and materials. Lab equipment for light microscopy includes confocal microscopes, fluorescence microscopes, zoom and stereo microscopes. Microscope slides and imaging reagents are available for visualizing samples, as well as various microscope stages and incubators for large or temperature-sensitive samples. Find the best light microscopes in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Protein ExpressionProtein expression is the utilization of cell machinery for the synthesis of proteins and has become a critical tool in biotherapeutic, genomic, and proteomic research. Produce recombinant proteins with expression vectors in combination with a host cell suitable for high-level protein expression. For production of toxic proteins, consider cell-free expression vectors. Create and monitor post-translational modifications with protein modification kits. Find the best protein expression products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Tissue ProcessingTissue processing involves preparing biological tissue samples for microscopic analysis, including fixation, embedding, and sectioning. This process is essential for histopathology and diagnostic applications. Explore tissue processing products in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.ImmunohistochemistryImmunohistochemistry (IHC) is a technique used to detect specific proteins in tissue samples by using antibodies that bind to target antigens. IHC is widely applied in pathology, immunology, hematology and cancer diagnostics. Explore IHC tools in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.CytometryCytometry is a broad term for the quantitative analysis of cells and cell systems. The most popular methods to study cytometry are flow cytometry and image cytometry.
Quantitating Protein Expression in Intact Tissue Sections for Cancer Research