TeloTAGGG™ Telomerase PCR ELISAPLUS
ROCHE/12013789001 - sufficient for ≤96 reactions, kit of 1 (16 components), suitable for enzyme immunoassay
Synonym: ELISA
Product Type: Chemical
manufacturer/tradename | Roche |
packaging | kit of 1 (16 components) |
Quality Level | 100 |
storage temp. | −20°C |
technique(s) | enzyme immunoassay: suitable |
usage | sufficient for ≤96 reactions |
Application: | The TeloTAGGG Telomerase PCR ELISAPLUS has been used to detect telomerase activity in urine, frozen sections of head and neck squamous cell carcinomas and tumor margin tissues. |
Features and Benefits: | • Ready-to-use master mixes: the kit contains all of the essential compounds as master mixes • Reliability: Positive control contained in the kit • Specificity: The use of hybridization probes ensures specific detection of the applied telomerase reaction product • Quantitative results: The assay concept allows semi-quantitative determination of telomerase levels |
General description: | The TeloTAGGG Telomerase PCR ELISAPLUS kit provides a way to perform a highly sensitive photometric enzyme immunoassay to detect telomerase activity, using nonradioactive techniques. The TeloTAGGG Telomerase PCR ELISAPLUS is designed for use in life science research studies in cell extracts from cell cultures and other biological research samples. The TeloTAGGG Telomerase PCR ELISAPLUS utilizes single-tube reaction mixtures, similar to the TeloTAGGG Telomerase PCR ELISA, but also contains an optimized internal standard and control template, which allows the TeloTAGGG Telomerase PCR ELISAPLUS to quantitatively measure the amount of telomerase reaction product. It uses the Telomeric Repeat Amplification Protocol (TRAP). |
Legal Information: | TeloTAGGG is a trademark of Roche |
Legal Information: | Manufactured under license from Geron Corporation. |
Other Notes: | For life science research only. Not for use in diagnostic procedures. |
Packaging: | 1 kit containing 16 components. |
Preparation Note: | Working solution: Solution 10: Washing Buffer, 1x Dilute an appropriate volume of Washing buffer, 10x concentrated (bottle 10) with autoclaved double-distilled water (1:10) and mix thoroughly. Approximately 5 ml of the diluted Washing buffer are needed for one reaction. Stable at 2 to 8 °C for 1 month. Solution 11: Anti-DIG HRP, bottle 11 stock solution Reconstitute the lyophilizate in 240 μl autoclaved double-distilled water. This results in an antibody conjugate concentration of 0.5 U/ml. Stable at 2 to 8 °C for 6 months. Do not freeze! Do not add sodium azide! Solution 11: Anti-DIG HRP, working solution To prepare the working solution, dilute an appropriate amount of the reconstituted anti-DIG-HRP (solution 11) with Conjugate dilution buffer (solution 12) to a final concentration of 10 mU/ml (e.g., 200 μl antibody solution and 9.8 ml of Conjugate dilution buffer). Prepare immediately before use. Do not store. Storage conditions (working solution): Washing buffer, 1x: Stable at 2 to 8 °C for 1 month. Anti-DIG HRP, stock solution: Stable at 2 to 8 °C for 6 months. Do not freeze. Do not add sodium azide. Anti-DIG HRP, working solution: Prepare immediately before use. Do not store. |
Principle: | The test principle can be divided into the following steps: Step 1: Elongation/amplification In the first step, telomerase adds telomeric repeats (TTAGGG) to the 3′-end of the biotin-labeled synthetic P1-TS primer. These elongation products, as well as the Internal standard (IS) included in the same reaction vessel, are amplified by PCR using the primers P1-TS and the anchor-primer P2. PCR products derived from telomerase-mediated elongation products in the first step contain the telomerase-specific six nucleotide increments, while the Internal standard (IS) generates a 216-bp PCR product. Step 2: Detection by ELISA The PCR products are split into two aliquots, denatured, and hybridized separately to digoxigenin-(DIG)-labeled detection probes that are specific for the telomeric repeats (P3-T) and for the Internal standard (IS) (P3-Std), respectively. The resulting products are immobilized via the biotin label to a streptavidin-coated microplate. Immobilized amplicons are then detected with an antibody against digoxigenin, which is conjugated to horseradish peroxidase (Anti-DIG-HRP) and the sensitive peroxidase substrate TMB. |
Specifications: | Assay time: Approximately 7 hours Measuring range: The linear measuring range of the kit is from 10 - 2,000 cells (in a model system using 293 cells) Sample material: Cell cultures and biological research samples Sensitivity: Detects <10 cell equivalents (in a model system, using 293 cells) |
Symbol | GHS05,GHS07 |
Signal word | Warning |
Hazard statements | H290 - H317 |
Precautionary statements | P261 - P280 - P333 + P313 - P362 + P364 - P390 - P501 |
RIDADR | UN 3316 9 |
WGK Germany | WGK 2 |
Flash Point(F) | does not flash |
Flash Point(C) | does not flash |
Storage Temp. | −20°C |
UNSPSC | 41116133 |
Components | Lysis Reagent; Reaction Mixture 2x concentrated; Internal Standard (IS); Control Template, low; Control Template, high; Water, nuclease-free; Denaturing Reagent; Hybridization Buffer T; Hybridization Buffer IS; Washing Buffer 10x concentrated; Anti-DIG-HRP antibody; Conjugate Dilution Buffer; TMB Substrate Solution; Stop Reagent; Microplate; Cover Foil |