Tyrosine Kinases

Human Tyrosine Kinases PCR Array The Human Tyrosine Kinases RT² Profiler PCR Array profiles the expression of 84 receptor and non-receptor tyrosine kinase genes. The protein tyrosine kinase superfamily includes roughly 60 receptor tyrosine kinases (RTKs) and about 30 intracellular tyrosine kinases. RTKs include an extracellular domain, a transmembrane domain, and a catalytic intracellular domain. Upon activation, RTKs dimerize and autophosphorylate their intracellular domain, initiating downstream signaling that often includes non-receptor tyrosine kinases. Non-receptor tyrosine kinases include a catalytic domain and a regulatory domain, which vary for each family. For example, the SRC-family kinase regulatory domain requires autophosphorylation for kinase domain activation, while most other intracellular tyrosine kinase families use different regulatory mechanisms. Tyrosine kinases are involved in many basic biological processes, such as growth, proliferation, and differentiation. These processes are commonly dysregulated during oncogenesis, often due to mutation of key tyrosine kinases or regulators. These oncogenic processes make the tyrosine kinase superfamily members attractive drug targets, and there are several chemotherapeutics targeting tyrosine kinases already on the market (e.g., imatinib mesylate). This array includes most RTKs and non-receptor tyrosine kinases. The results of this array can yield new insights into tyrosine kinase expression and regulation in an experimental model system. Using real-time PCR, research studies can easily and reliably analyze the expression of a focused panel of tyrosine kinase genes with this array. The RT² Profiler PCR Arrays are intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease. 96-well Plate, 384-well (4 × 96) Plate, and 100-well Disc formats are available. Available for cells, tissues, FFPE samples and small samples PAHS-152A-2 PAHS-152A-12 PAHS-152A-24 PAHS-152C-2 PAHS-152C-12 PAHS-152C-24 PAHS-152D-2 PAHS-152D-12 PAHS-152D-24 PAHS-152F-2 PAHS-152F-12 PAHS-152F-24 PAHS-152E-4 PAHS-152G-4 Protocol Guide

Functional Gene Grouping

How It Works

Manual & Resources

Reagents & Software

"If you have access to a real-time PCR instrument, you can use PCR Arrays" RT² Profiler™ PCR Arrays are the most reliable and sensitive gene expression profiling technology for analyzing a panel of genes in signal transduction pathways, biological process or disease related gene networks. The PCR Arrays can be used for research on cancer, immunology, stem cells, toxicology, biomarker discovery and validation, and phenotypic analysis of cells and transgenic animals. See the complete list of PCR Arrays. You can use any 96-well or 384-well real-time PCR instrument. See instrument-specific setup instructions. Click below for detailed information. How it Works Performance Data Application Data How it Works The PCR array is a set of optimized real-time PCR primer assays on 96-well or 384-well plates for pathway or disease focused genes as well as appropriate RNA quality controls. The PCR array performs gene expression analysis with real-time PCR sensitivity and the multi-gene profiling capability of a microarray. Simply mix your cDNA template with the appropriate ready-to-use PCR master mix, aliquot equal volumes to each well of the same plate, and then run the real-time PCR cycling program. (Download user manual) Figure 1: How PCR Arrays Work - Protocol Chart What it offers?      Guaranteed Performance* - ready-to-use for gene expression analysis      Time and cost saving - less than 30 min hands-on time for analyzing 84 genes      Ease of data analysis using our easy-to-use Excel-based data analysis template or web-based analysis tool Layout and Controls: The PCR Arrays are available in both 96- and 384-well plates and are used to monitor the expression of 84 genes related to a disease state or pathway plus five housekeeping genes. Controls are also included on each array for genomic DNA contamination, RNA quality, and general PCR performance You can easily perform a PCR Array experiment in your own laboratory, or send your samples to us and take advantage of our PCR Array Services. *: when using complete PCR array system. Performance Data Sensitivity: The complete PCR Array System yields a greater-than 85 percent present call with as little as 25 ng as much as 5 µg of total RNA from a pathway representing genes expressed at a lower level (inflammatory cytokines and receptors). Figure 2: PCR Arrays Let You See More Genes with Less RNA Different amounts of universal total RNA were characterized using the Human Inflammatory Cytokines and Receptors PCR Array, and the percentage of detectable genes was calculated for each RNA amount. As little as 25 ng total RNA yields greater than an 80% positive call, even for cytokines expressed at very low levels. Reproducibility The complete PCR Array System demonstrates strong correlations across technical replicates, lots, and instruments with average correlation coefficients > 0.99 insuring reliable detection of differences in expression between biological samples.   050825_1  050825_2  050825_3  050825_4 060111_1 0.993 0.989 0.995 0.992 060111_2 0.994 0.990 0.995 0.992 060111_3 0.992 0.990 0.993 0.992 060111_4 0.993 0.992 0.994 0.992 Figure 3: PCR Arrays Yield Highly Reproducible Results Four replicate sets of raw threshold data (1-4) obtained by two different scientists (A & B) at two different times (050825 & 060111) on Human Drug Metabolism RT² Profiler PCR Arrays are directly compared. The results demonstrate a high degree of correlation (R2 > 0.990). Specificity The complete PCR Array System, with high quality input RNA, is guaranteed to yield single bands of the predicted size without primer dimers or other secondary products thus providing the most accurate real-time PCR results possible. Figure 4: PCR Arrays Amplify A Single Gene-Specific Product in Every Reaction. Universal total RNA was characterized on the TGFβ / BMP Signaling Pathway PCR Array, followed by dissociation (melt) curve analysis. PCR Arrays specifically detect individual genes despite the expression of related gene family members in the same RNA sample. Application Data Cancer Research: To ascertain the oncogenic route that two different human breast tumors have taken, the relative expression level of cancer- and adhesion-related genes in normal and two different cancerous tissues were compared. Template cDNAs prepared from total RNA of normal human breast and two human breast tumors (BioChain Institute, Inc., 5.0 µg) were characterized in technical triplicates using the Human Cancer PathwayFinder™ PCR Array and the Human Extracellular Matrix & Adhesion Molecule PCR Array with the RT² SYBR Green / Fluorescein PCR master mix on the Bio-Rad iCycler®. Figure 5: ECM & Adhesion PCR Arrays Revealed Up- and Down-Regulated Genes in Breast Cancer Total RNA from normal human breast and a human breast tumor were characterized in technical triplicates, and the relative expression levels for each gene in the two samples are plotted against each other in the Scatter Plot. Genes encoding the matrix metallopeptidases (MMP3 & MMP9) and their inhibitors (TIMP3) are up-regulated, while genes encoding integrins (ITGB3 & ITGB4) are down-regulated, by at least three-fold (outside the silver field) in breast tumors relative to normal tissue. Toxicology Research: Rezulin (Troglitazone or "Tro" or "T"), a glitazone PPAR-gamma agonist, was approved for treatment of type 2 diabetes mellitus, but was withdrawn from the market due to idiosyncratic liver toxicity. Two similar drugs, Avandia (Rosiglitazone or "Rosi" or "R") and Actos (Pioglitazone or "Pio" or "P"), are considered to be safe treatments for the same condition. The expression profile of key drug metabolism genes should be different in cells treated with Rezulin versus those treated with Avandia and Actos. Hepatocellular carcinoma HepG2 cells were treated at 80% cell confluence with these three drugs (100 µM, Cayman Chemical) or a DMSO vehicle control for 24 h. RNA isolated using the ArrayGrade™ Total RNA Isolation Kit was used to characterize gene expression with the Human Drug Metabolism and Stress & Toxicity PathwayFinder™ RT² Profiler™ PCR Arrays and RT² SYBR Green / Fluorescein PCR master mix on the Bio-Rad iCycler®. Figure 6: Stress & Toxicity PathwayFinder™ PCR Array Uncovered Idiosyncratic Mechanisms of Action for Liver Toxicity Caused by 3 PPARγ Agonists. RNA from HepG2 cells treated with three different glitazone PPARγ agonists for type 2 diabetes mellitus was characterized, and the results were compared to that of a vehicle (DMSO) control. A withdrawn drug with idiosyncratic liver toxicity (Rezulin) induces very different changes in the expression of stress-related genes than two safer drugs still on the market (Avandia and Actos). Back to Top

User Manual RT² Profiler™ PCR Array User Manual (PDF) White Paper PCR Array: A Simple and Quantitative Method for Gene Expression Profiling (PDF) PCR Array Application Examples: Toxicology, Oncology, and Immunology Research (PDF) Data Analysis Free PCR Array Data Analysis Software (Web & Excel based) Instrument Setup Instructions Materials & Equipment Required for our PCR products PCR Array Instrument Setup Instructions & Protocol Files FAQ FAQ about Real-Time PCR and Troubleshooting for our PCR products Web Seminars Attend a live on-line seminar hosted by an Applications Scientist about PCR Technical Brochures PCR Array Brochure (PDF) Primer Assay Brochure (PDF) SYBR Green Master Mix Brochure (PDF) Powerpoint Presentations PCR Technology Overview Presentations Citations & Publications Literature Citations using our PCR product Pathway Central Pathway reviews and presentation-ready pathway maps shRNA Plasmids Pathway related gene knockout - guaranteed > 70% knockdown Individual Primers Validated and Certified qPCR Primers for the genes on this array Back to Top