S7710 | TRAPEZE® RT Telomerase Detection Kit

S7710
1 kit  The kit provides enough reagents to perform 224 TRAPEZE® RT reactions.
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      Overview

      Replacement Information

      Key Specifications Table

      Description
      Catalogue NumberS7710
      Brand Family Chemicon®
      Trade Name
      • TRAPeze
      • Chemicon
      DescriptionTRAPEZE® RT Telomerase Detection Kit
      OverviewThe TRAPeze® RT Telomerase Detection Kit is a highly sensitive in vitro assay for the fluorometric detection and real time quantification of telomerase activity. It incorporates refinements to the original TRAPeze assay and adds the ability to quantitate telomerase activity using fluorescence energy transfer (ET) primers.
      Alternate Names
      • TRAP Assay
      Background InformationTelomeres are specific structures found at the end of chromosomes in eukaryotes. In human chromosomes, the telomeres consist of thousands of copies of 6 base repeats (TTAGGG)(1-3). It has been suggested that telomeres protect chromosome ends since damaged chromosomes lacking telomeres undergo fusion, rearrangement and translocation (2). In somatic cells, telomere length is progressively shortened with each cell division both in vivo and in vitro (4-7) due to the inability of the DNA polymerase complex to synthesize the very 5' end of the lagging strand (8,9).

      Telomerase is a ribonucleoprotein that synthesizes and directs the telomeric repeats onto the 3' end of existing telomeres using its RNA component as a template (10-14). Telomerase activity has been shown to be specifically expressed in immortal cells, cancer and germ cells (15,16) where it compensates for telomere shortening during DNA replication and thus stabilizes telomere length (7,17). These observations have led to a hypothesis that telomere length may function as a "mitotic clock" to sense the number of cell divisions and eventually signal replicative senescence or programmed cell death when a critical telomere length is achieved. Therefore, expression of telomerase activity in cancer cells may be a necessary and essential step for tumor development and progression (16,18-20). The causal relationship between expression of telomerase and telomere length stabilization and the extension of the life span of the human cell has recently been reported (21).

      The development of a sensitive and efficient PCR-based telomerase activity detection method, TRAP (Telomeric Repeat Amplification Protocol) (15, 22), has made possible large scale surveys of telomerase activity in human cells and tissues (15, 23-29). To date, telomerase activity has been detected in over 85% of all tumors tested spanning more than 20 different types of cancers (30-31).

      The TRAPEZE® RT Telomerase Detection Kit is a highly sensitive in vitro assay for the fluorometric detection and real time quantification of telomerase activity. It incorporates refinements to the original TRAP assay that were first introduced in the gel-based TRAPEZE® Telomerase Detection Kit (Cat. No. S7700) and adds the ability to quantitate telomerase activity using fluorescence energy transfer (ET) primers similar to the TRAPEZE® XL Telomerase Detection Kit (Cat. No. S7707). As in the original TRAPEZE® Kit, primer sequence modifications that reduce amplification artifacts and PCR controls for standard curve generation are included. In addition, both the TRAPEZE® RT and XL Kits use fluorescence energy transfer (ET) primers to generate fluorescently labeled TRAP products which permit nonisotopic, quantitative analysis of telomerase activity.

      The unique design of these ET primers (Amplifluor® primers) allows detection and quantification of telomerase activity by directly measuring real time fluorescence emission in the reaction vessels. Since Amplifluor® primers will fluoresce only upon incorporation into the TRAP products, post-PCR sample manipulations such as electrophoretic gel or ELISA analyses are eliminated, thereby reducing the the risk of carry-over contamination. Quantitative analysis is not compromised when detection is performed in a high-throughput 96-well format unlike platforms utilizing a qualitative ELISA. Additionaly, an additional stand alone control is provided separately to assess PCR inhibitors that may be present in experimental samples. (Please see product insert for references).
      Materials Required but Not DeliveredEquipment and Supplies
      1. Real time fluorescence capable thermocycler with FAM detection (i.e. ABI Prism®7700/7900, iCycler®, MJ Opticon™, Stratagene MX® 4000, etc.) (See Sec. V. Appendix, Excitation and Emission Filters in product insert)
      2. Optically clear 96-well plates and caps for real-time PCR amplification and detection
      3. If analyzing tissues, homogenization equipment as described in Sec. III. Protocol, Extract Preparation 4. Aerosol resistant pipette tips (RNase-free)

      Reagents
      1. Antibody-mediated, hot start Taq polymerase Note: Titanium™ Taq (BD Clontech®) is recommended for use with the TRAPEZE® RT Telomerase Detection System. Other antibody hot start enzymes may be suitable for use with this assay with optimization. Non-hot start and chemically modified hot start enzymes are not recommended for use with Amplifluor®.
      2. PBS (Mg2+- and Ca2+-free)
      3. Reagents for protein concentration measurement
      4. RNase inhibitor (for extract preparation from tissues)
      5. Bovuminar® Bovine Serum Albumin
      References
      Product Information
      Components
      • CHAPS Lysis Buffer (13.5mL)
      • 5X TRAPEZE® RT Reaction Mix (1.12mL)
      • 5X TRAPEZE® Control Reaction Mix (1.12mL)
      • PCR - Grade Water (8.2mL)
      • TSR8* (quantitation control template) (45 μL)
      • TSK* (pcr inhibition/normalization control) (45 μL)
      • Control Cell Pellet (Telomerase positive cells; 106 cells)
      • * Caution - refer to Sec. II. Kit Components, Precautions in product insert.
      Applications
      ApplicationA highly sensitive in vitro assay for the fluorometric detection & real time quantification of telomerase activity in cells.
      Biological Information
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage Conditions1. CHAPS Lysis Buffer - 15°C to -25°C
      2. 5X TRAPEZE® RT Reaction Mix -15°C to -25°C
      3. 5X TRAPEZE® Control Reaction Mix 2°C to 8°C
      4. PCR - Grade Water - 15°C to -25°C
      5. TSR8 -15°C to -25°C
      6. TSK -15°C to -25°C
      7. Control Cell Pellet -75°C to -85°C
      Packaging Information
      Material Size1 kit
      Material PackageThe kit provides enough reagents to perform 224 TRAPEZE® RT reactions.
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      SDS

      Title

      Safety Data Sheet (SDS) 

      References

      Reference overviewPub Med ID
      Estrogen and telomerase in human peripheral blood mononuclear cells.
      Ann L Benko,Nancy J Olsen,William J Kovacs
      Molecular and cellular endocrinology  364  2012

      Show Abstract
      22954679 22954679
      Abeta peptides accelerate the senescence of endothelial cells in vitro and in vivo, impairing angiogenesis.
      Sandra Donnini,Raffaella Solito,Elisa Cetti,Federico Corti,Antonio Giachetti,Silvia Carra,Monica Beltrame,Franco Cotelli,Marina Ziche
      FASEB journal : official publication of the Federation of American Societies for Experimental Biology  24  2010

      Show Abstract
      20207941 20207941
      Cullin 5 regulates cortical layering by modulating the speed and duration of Dab1-dependent neuronal migration.
      Simó S, Jossin Y, Cooper JA
      J Neurosci  30  5668-76.  2010

      Show Abstract
      20410119 20410119

      Technical Info

      Title
      Millipore Tools for Characterizing Induced Pluripotent Stem Cells

      User Guides

      Title
      TRAPeze® Kit RT Telomerase Detection Kit