S7710
Kit de détection de la télomérase TRAPeze™ RT
A highly sensitive in vitro assay for the fluorometric detection & real time quantification of telomerase activity in cells.
Synonyme(s) :
TRAP Assay
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About This Item
Code UNSPSC :
12161503
eCl@ss :
32161000
Nomenclature NACRES :
NA.32
Produits recommandés
Niveau de qualité
Fabricant/nom de marque
Chemicon®
TRAPeze™
Application(s)
genomic analysis
Conditions d'expédition
dry ice
Description générale
Le kit de détection de la télomérase TRAPeze™ RT est un dosage in vitro extrêmement sensible pour la détection fluorimétrique et la quantification en temps réel de l'activité télomérase. Il bénéficie d'améliorations par rapport au dosage TRAPeze™ d'origine, auxquelles s'ajoute la possibilité de quantifier l'activité télomérase avec des amorces à transfert d'énergie de fluorescence.
Telomeres 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).
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).
Conditionnement
Ce kit contient suffisamment de réactifs pour effectuer 224 réactions TRAPeze™ RT.
Composants
Tampon de lyse CHAPS (13,5 ml)
Mélange réactionnel TRAPeze™ RT concentré 5 × (1,12 ml)
Mélange réactionnel témoin TRAPeze™ concentré 5 × (1,12 ml)
Eau de qualité PCR (8,2 ml)
TSR8* (matrice témoin de quantification) (45 μl)
TSK* (témoin d'inhibition/normalisation de la PCR) (45 μl)
Culot de cellules témoin (cellules positives pour la télomérase ; 106 cellules)
* Attention : voir le chapitre II ("Kit Components, Precautions") de la notice.
Mélange réactionnel TRAPeze™ RT concentré 5 × (1,12 ml)
Mélange réactionnel témoin TRAPeze™ concentré 5 × (1,12 ml)
Eau de qualité PCR (8,2 ml)
TSR8* (matrice témoin de quantification) (45 μl)
TSK* (témoin d'inhibition/normalisation de la PCR) (45 μl)
Culot de cellules témoin (cellules positives pour la télomérase ; 106 cellules)
* Attention : voir le chapitre II ("Kit Components, Precautions") de la notice.
Stockage et stabilité
1. Tampon de lyse CHAPS : entre -15 °C et -25 °C
2. Mélange réactionnel TRAPeze™ RT concentré 5 × : entre -15 °C et -25 °C
3. Mélange réactionnel témoin TRAPeze™ concentré 5 × : entre 2 °C et 8 °C
4. Eau de qualité PCR : entre -15 °C et -25 °C
5. TSR8 : entre -15 °C et -25 °C
6. TSK : entre -15 °C et -25 °C
7. Culot de cellules témoin : entre -75 °C et -85 °C
2. Mélange réactionnel TRAPeze™ RT concentré 5 × : entre -15 °C et -25 °C
3. Mélange réactionnel témoin TRAPeze™ concentré 5 × : entre 2 °C et 8 °C
4. Eau de qualité PCR : entre -15 °C et -25 °C
5. TSR8 : entre -15 °C et -25 °C
6. TSK : entre -15 °C et -25 °C
7. Culot de cellules témoin : entre -75 °C et -85 °C
Informations légales
ABI PRISM is a registered trademark of Applera Corporation or its subsidiaries in the US and/or certain other countries
Amplifluor is a registered trademark of Merck KGaA, Darmstadt, Germany
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
Opticon is a trademark of Bio-Rad Laboratories, Inc.
TRAPEZE is a trademark of Merck KGaA, Darmstadt, Germany
iCycler is a registered trademark of Bio-Rad
Mention d'avertissement
Warning
Mentions de danger
Conseils de prudence
Classification des risques
Aquatic Chronic 3 - Met. Corr. 1
Code de la classe de stockage
8B - Non-combustible corrosive hazardous materials
Point d'éclair (°F)
Not applicable
Point d'éclair (°C)
Not applicable
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