细胞死亡通路发现者PCR芯片 Cell Death Path

“英拜为您实验加速”
技术服务网址：http://www.yingbio.com/
服务热线：400-696-6643、 18019265738
邮箱：daihp@yingbio.com 、 huizhang1228@foxmail.com

Cell Death PathwayFinder PCR Array

细胞死亡通路发现者PCR芯片

Product	Species	Technology	Cat. No.
Cell Death PathwayFinder PCR Array	Human	Gene Expression	PAHS-212Z
Cell Death PathwayFinder PCR Array	Mouse	Gene Expression	PAMM-212Z
Cell Death PathwayFinder PCR Array	Rat	Gene Expression	PARN-212Z

The Human Cell Death PathwayFinder RT² Profiler PCR Array profiles the expression of 84 key genes important for the central mechanisms of cellular death: apoptosis, autophagy, and necrosis. Apoptosis, or programmed cell death, results in controlled cell shrinkage and fragmentation via the action of caspases, as well as an anti-inflammatory cytokine release. In contrast, necrosis signals via RIPK1 (RIP1), leading to cell swelling, lysis, and a pro-inflammatory cytokine release. Autophagy destroys the cell's damaged proteins and organelles via an intracellular catabolic process in the lysosome. Multiple cellular processes require the removal of specific cells by a controlled cell-death program. For example, tissue remodeling activates apoptosis, whereas energy metabolism and growth regulation responses rely on autophagy. Developmental processes often activate apoptosis, while bodily injuries or infection more commonly induce necrosis. The molecular mechanisms behind these cell death pathways overlap and more than one form of cell death occur simultaneously during some cellular functions. Apoptosis and necrosis both signal through the death domain receptors FAS, TNFRSF1A (TNFR1), and TNFRSF10A (TRAIL-R), while autophagy and apoptosis share BCL2 family members as key players. The results of this array can yield insights into which central cell death mechanism(s) drive normal biological or pathophysiological processes. Using real-time PCR, research studies can easily and reliably analyze the expression of a focused panel of genes involved in cellular death pathways 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.
细胞死亡通路发现者PCR芯片可用于研究参与细胞死亡的中心机制：凋亡、自噬、坏死的84个关键基因的表达。细胞凋亡或程序性细胞死亡有两种情况：半胱氨酸酶和抗炎细胞因子释放，进而导致细胞收缩和破碎；坏死信号通过RIPK1（RIP1）的释放，则导致细胞膨胀和溶解。自噬可以通过细胞内的溶酶体清除细胞中受损的蛋白和细胞器。细胞程序性死亡在生物学过程中时常发生，例如组织重塑会激活细胞凋亡，能量代谢和生长调节也离不开自噬；发育过程会激活细胞凋亡，身体受伤或感染则伴随坏死。凋亡和坏死与FAS、TNFRSF1A（TNFR1）、TNFRSF10A有关；自噬和凋亡则与BCL2家庭有关。使用实时定量PCR，研究者可以很简单和可靠地分析参与细胞死亡通路的基因表达。通过实时定量PCR的方法，研究者即能够利用该芯片简单可靠地同时检测细胞死亡通路相关基因的表达，研究与之相关的正常生理或病理过程的驱动机制
Apoptosis:
Pro-Apoptotic: ABL1, APAF1, ATP6V1G2, BAX, BCL2L11, BIRC2 (c-IAP2), CASP1 (ICE), CASP3, CASP6, CASP7, CASP9, CD40 (TNFRSF5), CD40LG (TNFSF5), CFLAR (CASPER), CYLD, DFFA, FAS (TNFRSF6), FASLG (TNFSF6), GADD45A, NOL3, SPATA2, SYCP2, TNF, TNFRSF1A, TNFRSF10A (TRAIL-R), TP53.
Anti-Apoptotic: AKT1, BCL2, BCL2A1 (Bfl-1/A1), BCL2L1 (BCL-X), BIRC3 (c-IAP1), CASP2, IGF1R, MCL1, TNFRSF11B, TRAF2, XIAP.
Autophagy: AKT1, APP, ATG12, ATG16L1, ATG3, ATG5, ATG7, BAX, BCL2, BCL2L1 (BCL-X), BECN1, CASP3, CTSB, CTSS, ESR1 (ERa), FAS (TNFRSF6), GAA, HTT, IFNG, IGF1, INS, IRGM, MAP1LC3A, MAPK8 (JNK1), NFKB1, PIK3C3 (VPS34), RPS6KB1, SNCA, SQSTM1, TNF, TP53, ULK1.
Necrosis: ATP6V1G2, BMF, C1orf159, CCDC103, COMMD4, CYLD, DEFB1, DENND4A, DPYSL4, EIF5B, FOXI1, GALNT5, GRB2, HSPBAP1, JPH3, KCNIP1, MAG, OR10J3, PARP1 (ADPRT1), PARP2, PVR, RAB25, S100A7A, SPATA2, SYCP2, TMEM57, TNFRSF1A, TXNL4B.

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).