TeloTAGGG™ Telomerase PCR ELISAPLUS 12013789001

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).
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

Packaging
1 kit containing 16 components.
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)
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.
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.
For life science research only. Not for use in diagnostic procedures.
Manufactured under license from Geron Corporation.
TeloTAGGG is a trademark of Roche

Quality Level	100,
usage	sufficient for 96 reactions
Торговая марка	Roche
shipped in	dry ice
storage temp.	20°C

pictograms	GHS05,GHS07
signalword	Warning
hcodes	H290 - H317
pcodes	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