EpiTect® Methyl II Signature PCR Array (22) Human Notch Signaling Pathway Cat. no. 335212 EAHS-611ZA For DNA methylation analysis using MethylScreen™ technology The Human Notch Signaling Pathway EpiTect Methyl II Signature PCR Array profiles the promoter methylation status of a panel of 22 genes central to Notch signal transduction. The Notch signaling pathway regulates the expression of genes mediating cell-cell communication and developmental processes. DNA methylation status changes on key regulatory genes during development affect their ability to be induced or repressed by Notch and other signaling pathways. Profiling research samples containing genomic DNA from cellular or fresh tissue with this array may help to correlate CpG island methylation status with biological phenotypes such as developmental stages. This array contains key ligands, cofactors, receptors, downstream signaling effectors, and target genes for Notch signaling. The results may also help provide further insights into the molecular mechanisms and biological pathways behind the role of Notch signaling during cellular communication, differentiation, and development. With a simple restriction enzyme digestion and real-time PCR, research studies can analyze the promoter methylation status of 22 different genes central to Notch signal transduction with this DNA methylation PCR array. The EpiTect Methyl II PCR Arrays use MethylScreen™ technology provided under license from Orion Genomics, LLC. For further details, consult the EpiTect Methyl II PCR Array Handbook. Shipping and storage EpiTect Methyl II Signature PCR Arrays are shipped at room temperature (15–25°C), on dry ice, or on blue ice depending on the destination and accompanying products. They should be stored at –20°C. Ensure that you have the correct EpiTect Methyl II Signature PCR Array format for your instrument before starting the experiment. Note: Open the package and store the products appropriately immediately on receipt. Sample & Assay Technologies Contents Product Contents Cat. no. EpiTect Methyl II Signature PCR 2, 12, or 24 x 96-well plates, Optical Thin-Wall Varies Array (22) Format A 8-Cap Strips; for use with the following real-time cyclers: Applied Biosystems® models 5700, 7000, 7300, 7500, 7700, 7900HT, ViiA™ 7 (96-well block); Bio-Rad® models iCycler®, iQ™5, MyiQ™, MyiQ2; Bio-Rad/MJ Research Chromo4™; Eppendorf® MasterCycler® ep realplex models 2, 2s, 4, 4s; Stratagene® models Mx3005P®, Mx3000P ® EpiTect Methyl II Signature PCR 2, 12, or 24 x 96-well plates, Optical Adhesive Film; Varies Array (22) Format C for use with the following real-time cyclers: Applied Biosystems models 7500 (Fast block), 7900HT (Fast block), StepOnePlus™, ViiA 7 (Fast block) EpiTect Methyl II Signature PCR 2, 12, or 24 x 96-well plates, Optical Thin-Wall Varies Array (22) Format D 8-Cap Strips; for use with the following real-time cyclers: Bio-Rad CFX96™; Bio-Rad/MJ Research models DNA Engine Opticon®, DNA Engine Opticon 2; Stratagene Mx4000® EpiTect Methyl II Signature PCR 4 x 384-well plates, 384EZLoad Covers, Optical Varies Array (22) Format E Adhesive Film; for use with the following real-time cyclers: Applied Biosystems models 7900HT (384-well block), ViiA 7 (384-well block); Bio-Rad CFX384™ EpiTect Methyl II Signature PCR 2, 12, or 24 x 96-well plates, Optical Adhesive Film; Varies Array (22) Format F for use with the following real-time cyclers: Roche® LightCycler® 480 (96-well block) EpiTect Methyl II Signature PCR 4 x 384-well plates, 384EZLoad Covers, Optical Varies Array (22) Format G Adhesive Film; for use with the following real-time cyclers: Roche: LightCycler 480 (384-well block) Array layout 1 2 3 4 5 6 7 8 9 10 11 12 A ADAM17 CCND1 CREBBP CTBP1 DLL1 DLL3 DVL3 ERBB2 HDAC2 HES1 JAG1 KAT2A B LFNG MAML1 NCOR2 NCSTN NFKB2 NOTCH1 NUMBL PSENEN RBPJL SNW1 EP_SEC EP_DEC C ADAM17 CCND1 CREBBP CTBP1 DLL1 DLL3 DVL3 ERBB2 HDAC2 HES1 JAG1 KAT2A D LFNG MAML1 NCOR2 NCSTN NFKB2 NOTCH1 NUMBL PSENEN RBPJL SNW1 EP_SEC EP_DEC E ADAM17 CCND1 CREBBP CTBP1 DLL1 DLL3 DVL3 ERBB2 HDAC2 HES1 JAG1 KAT2A F LFNG MAML1 NCOR2 NCSTN NFKB2 NOTCH1 NUMBL PSENEN RBPJL SNW1 EP_SEC EP_DEC G ADAM17 CCND1 CREBBP CTBP1 DLL1 DLL3 DVL3 ERBB2 HDAC2 HES1 JAG1 KAT2A H LFNG MAML1 NCOR2 NCSTN NFKB2 NOTCH1 NUMBL PSENEN RBPJL SNW1 EP_SEC EP_DEC Gene table GenBank OR Gene Position Unigene miRNA Symbol OR Description Gene Name Accession miRNA ID ADAM18, CD156B, CSVP, A01, C01, E01, G01 Hs.404914 NM_003183 ADAM17 ADAM metallopeptidase domain 17 MGC71942, TACE BCL1, D11S287E, PRAD1, A02, C02, E02, G02 Hs.523852 NM_053056 CCND1 Cyclin D1 U21B31 A03, C03, E03, G03 Hs.459759 NM_004380 CREBBP CREB binding protein CBP, KAT3A, RSTS A04, C04, E04, G04 Hs.208597 NM_001328 CTBP1 C-terminal binding protein 1 BARS, MGC104684 A05, C05, E05, G05 Hs.379912 NM_005618 DLL1 Delta-like 1 (Drosophila) DELTA1, DL1, Delta A06, C06, E06, G06 Hs.127792 NM_016941 DLL3 Delta-like 3 (Drosophila) SCDO1 Dishevelled, dsh homolog 3 A07, C07, E07, G07 Hs.388116 NM_004423 DVL3 KIAA0208 (Drosophila) V-erb-b2 erythroblastic leukemia viral CD340, HER-2, HER-2, neu, oncogene homolog 2, A08, C08, E08, G08 Hs.446352 NM_001005862 ERBB2 HER2, MLN 19, NEU, NGL, neuro/glioblastoma derived oncogene TKR1 homolog (avian) A09, C09, E09, G09 Hs.3352 NM_001527 HDAC2 Histone deacetylase 2 HD2, RPD3, YAF1 Hairy and enhancer of split 1, FLJ20408, HES-1, HHL, HRY, A10, C10, E10, G10 Hs.250666 NM_005524 HES1 (Drosophila) bHLHb39 AGS, AHD, AWS, CD339, A11, C11, E11, G11 Hs.728907 NM_000214 JAG1 Jagged 1 HJ1, JAGL1, MGC104644 GCN5, GCN5L2, A12, C12, E12, G12 Hs.463045 NM_021078 KAT2A K(lysine) acetyltransferase 2A MGC102791, PCAF-b, hGCN5 LFNG O-fucosylpeptide B01, D01, F01, H01 Hs.159142 NM_001040167 LFNG 3-beta-N-acetylglucosaminyltransferas SCDO3 e FLJ53540, KIAA0200, B02, D02, F02, H02 Hs.631951 NM_014757 MAML1 Mastermind-like 1 (Drosophila) Mam-1, Mam1 CTG26, FLJ25011, N-CoR2, SMAP270, SMRT, SMRTE, B03, D03, F03, H03 Hs.137510 NM_006312 NCOR2 Nuclear receptor corepressor 2 SMRTE-tau, TNRC14, TRAC, TRAC-1, TRAC1 B04, D04, F04, H04 Hs.517249 NM_015331 NCSTN Nicastrin APH2, KIAA0253 Nuclear factor of kappa light B05, D05, F05, H05 Hs.73090 NM_002502 NFKB2 polypeptide gene enhancer in B-cells 2 LYT-10, LYT10, p52 (p49/p100) B06, D06, F06, H06 Hs.495473 NM_017617 NOTCH1 Notch 1 TAN1, hN1 CAG3A, CTG3a, NBL, B07, D07, F07, H07 Hs.326953 NM_004756 NUMBL Numb homolog (Drosophila)-like NUMB-R, NUMBLIKE, NUMBR, TNRC23 Presenilin enhancer 2 homolog (C. B08, D08, F08, H08 Hs.534465 NM_172341 PSENEN MSTP064, PEN-2, PEN2 elegans) Recombination signal binding protein B09, D09, F09, H09 Hs.248217 NM_014276 RBPJL for immunoglobulin kappa J RBP-L, RBPSUHL, SUH, SUHL region-like Bx42, MGC119379, B10, D10, F10, H10 Hs.445498 NM_012245 SNW1 SNW domain containing 1 NCOA-62, PRPF45, Prp45, GenBank OR Gene Position Unigene miRNA Symbol OR Description Gene Name Accession miRNA ID SKIIP, SKIP B11, D11, F11, H11 N/A SA_00148 EP_SEC SEC-sensitive enzyme control assay SEC B12, D12, F12, H12 N/A SA_00149 EP_DEC DEC-dependent enzyme control assay DEC Related products Product Contents Cat. no. EpiTect Methyl II DNA Reagents for the cleavage of methylated and 335452 Restriction Kit (12) unmethylated DNA for processing up to 12 DNA samples; 5x Restriction Digestion Buffer, Methylation-sensitive Enzyme A, Methylation-dependent Enzyme B RT² SYBR® Green qPCR For 2 x 96 assays in 96-well plates; suitable for use 330500 Mastermixes(2)* with real-time cyclers that do not require a reference dye, including: Bio-Rad models CFX96, CFX384; Bio-Rad/MJ Research models Chromo4, DNA Engine Opticon 2; Roche LightCycler 480 (96-well and 384-well); all other cyclers RT² SYBR Green ROX™ qPCR For 2 x 96 assays in 96-well plates; suitable for use 330520 Mastermixes(2)* with the following real-time cyclers: Applied Biosystems models 5700, 7000, 7300, 7500 [Standard and Fast], 7700, 7900HT 96-well block [Standard and Fast] and 384-well block, StepOnePlus; Eppendorf Mastercycler ep realplex models 2, 2S, 4, 4S; Stratagene models Mx3000P, Mx3005P, Mx4000 RT² SYBR Green Fluor qPCR For 2 x 96 assays in 96-well plates; suitable for use 330510 Mastermixes(2)* with the following real-time cyclers: Bio-Rad models iCycler, iQ5, MyiQ, MyiQ2 Accessories EpiTect Methyl II Custom PCR For methylation analysis of customer-selected genes 335112 Array in a 96-well or 384-well plate format EpiTect Methyl II Complete For methylation analysis of 94 genes in a 96-well or 335222 PCR Array (94) 384-well plate format EpiTect Methyl II PCR Assay Laboratory-tested forward and reverse primers for 335002 (200) 200 x 25 µl reactions; 25 µl per primer; total volume: 200 µl RT² PCR Array Loading 12 x 5 ml capacity, irradiation sterilized reservoirs 338162 Reservoir for convenient sample loading on PCR arrays * Larger kit sizes available; please inquire. EpiTect Methyl II Signature PCR Array products are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease. QIAGEN reserves the right to occasionally redesign individual assays on the EpiTect Methyl II PCR Arrays for improved performance. This revision history can be obtained by contacting technical support and providing the batch numbers from your arrays. For up-to-date licensing information and product-specific disclaimers, see the respective QIAGEN kit handbook or user manual. QIAGEN kit handbooks and user manuals are available at www.qiagen. com or can be requested from QIAGEN Technical Services or your local distributor. Purchaser agrees that use of this product and data therefrom is limited solely to the purchaser and for only the purchaser's own internal molecular biology research applications ("Permitted Use"), and shall not be re-sold or used for any other purposes (all of which are expressly prohibited), including without limitation diagnostic purposes, uses that could require regulatory approval for diagnostics from an agency of any government or regulatory entity anywhere in the world, diagnosis, prevention, or treatment of disease, and the right to perform commercial services of any kind, including without limitation, reporting the results of purchaser's activities, including without limitation, for a fee or other commercial consideration.