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Product Description
SHENTEK® Telomerase Assay Kit uses a Real-time Quantitative TRAP method (RQ-TRAP) implemented as a dual-fluorescence qPCR system. The assay includes an internal reference gene (IC, CY5) to control for inhibitors and false negatives, and uses TSR8 as a quantitative reference standard for accurate telomerase activity quantification (TPG Units). The kit is provided as a closed-cap qPCR assay (no gel electrophoresis or ELISA required) to improve throughput and reduce contamination risk. The kit includes reagents for cell lysis, RNase inhibition, TRAP qPCR reagents, TSR8 standard and telomerase positive cell controls and is designed for measuring telomerase activity in cell extracts.
Technical Specifications
| Parameter | Details |
|---|---|
| Assay format | Dual-fluorescence real-time quantitative TRAP (RQ-TRAP) qPCR |
| Kit throughput | Reagents for 200 reactions |
| Reaction volume | 25 μL total per reaction (23 μL qPCR MIX + 2 μL template) |
| Template volume per reaction | 2 μL |
| qPCR MIX per reaction | 2× TRAP qPCR Reaction Buffer 12.5 μL; TRAP qPCR Reaction Enzyme 1 μL; TRAP Primer&Control MIX 3 μL; TRAP qPCR Reaction Replenisher 6.5 μL |
| qPCR program (ABI7500 example) | Activation: 30 °C 30:00 (1 cycle); Activation: 95 °C 02:00 (1 cycle); 35 cycles of 95 °C 00:15 (denaturation), 50 °C 01:00 (anneal), 68 °C 00:30 (extension, fluorescence read) |
| Standard curve (TSR8) | ST1: 20 amol/μL (40000 TPG Units); ST2: 2 amol/μL (4000 TPG Units); ST3: 0.2 amol/μL (400 TPG Units); ST4: 0.02 amol/μL (40 TPG Units); ST5: 0.002 amol/μL (4 TPG Units); ST6: 0.0002 amol/μL (0.4 TPG Units). Note: ST0 = 200 amol/μL (400000 TPG Units). 1 amol TSR8 = 1000 TPG Units. |
| Dynamic / linear range | 0.4 to 400,000 TPG Units (based on ST6 - ST0 standard series) |
| Acceptance criteria for standard curve | Amplification efficiency 83.3%–110%; R² ≥ 0.990 |
| Detection limit (lowest standard) | 0.4 TPG Units (corresponding to 0.0002 amol/μL TSR8, ST6) — use instrument and software settings to confirm detection limit in your system |
| Sample input recommendations | Start from freshly harvested 10^6 cells (viability ≥80%) for extraction; for high-activity samples a 10-fold dilution or extraction from 10^5 cells recommended; for low-activity samples test undiluted extracts. Heat-inactivated extract (85 °C, 10 min) is required as negative control. |
| Telomerase positive control | Telomerase positive cells Pellet (NNA060) supplied; recommended processing described in protocol |
| Storage stability (kit) | Kit components can be stored under recommended conditions for up to 18 months; recommended storage –20 °C (short-term below –30 °C acceptable per USP <659>) |
| Instrument compatibility (examples) | ABI 7500, SHENTEK-96S, LightCycler 480 II (others may be used with appropriate programming/software adjustments) |
Features
- Dual-fluorescence qPCR (RQ-TRAP): Combines telomeric repeat amplification (TRAP) with real-time dual-channel detection (FAM for telomerase product, CY5 for internal control) for quantitative measurement and inhibition monitoring.
- Internal reference (IC): Internal reference gene (CY5) included to detect sample inhibition and reduce false negatives.
- Quantitative reference standard (TSR8): TSR8 ssDNA standard with defined amol/μL to convert qPCR signal to TPG Units for absolute quantification.
- Closed-cap workflow: No post-PCR gel electrophoresis or ELISA steps required, reducing hands-on time and contamination risk.
- High sensitivity and wide dynamic range: Standard curve covers 0.4 to 400,000 TPG Units (ST6–ST0), enabling detection from very low to high telomerase activities.
Applications
- Quantitative measurement of telomerase activity in cultured cells
- Comparative telomerase activity analysis in tumor cell lines, immortalized cells and stem cells (e.g., hiPSCs)
- Telomerase research in primary cells and mesenchymal stem cells (hMSCs)
- Method development and validation of telomerase activity assays in research settings
Kit Contents
Cell Lysis Buffer (NND058) — 1.5 mL × 5 tubes–20 °C
RNase inhibitor (NND060) — 50 μL × 1 tube–20 °C
Telomerase positive cells Pellet (NNA060) — 10^5 cells × 2 tubes–20 °C
2× TRAP qPCR Reaction Buffer (NNB022) — 650 μL × 4 tubes–20 °C, protect from light
TRAP qPCR Reaction Enzyme (NNC108) — 100 μL × 2 tubes–20 °C, protect from light
TSR8 (NNA059) — 50 μL × 1 tube (ssDNA quantitative reference)–20 °C, protect from light
TRAP Primer & Control MIX (NNC109) — 300 μL × 2 tubes–20 °C, protect from light
TRAP qPCR Reaction Replenisher (NND059) — 650 μL × 2 tubes–20 °C, protect from light
* Total: 200 reactions | Method: qPCR (Real-time Quantitative TRAP, RQ-TRAP)
Attention
• For research use only — read Material Safety Data Sheets (MSDS) and follow laboratory safety procedures; wear PPE.
• Maintain physically separated areas for sample preparation, positive area, negative area and PCR amplification to avoid cross-contamination.
• Use designated pipettes and aerosol-resistant RNase-free tips for TSR8 and standards; do not reuse those pipettes in other steps.
• Prepare Cell Lysis Buffer Working Solution by adding 5 μL RNase inhibitor to 1 mL Cell Lysis Buffer and keep on ice; use on ice to preserve telomerase activity.
• Heat inactivate each sample extract at 85 °C for 10 minutes to generate negative control (required).
• Avoid repeated freeze–thaw of cell pellets and extracts; cell pellets may be stored at –70 °C up to one year; extracts aliquoted and stored at –70 °C up to 6 months.
• Clean PCR tube racks with 10% bleach and UV irradiation after each use; prevent TSR8 and PCR product contamination of pre-PCR areas.
• Follow instrument-specific settings and analysis parameters; default threshold recommended is 0.1 and use Manual Ct with Automatic Baseline for analysis (as example).
Quality Management & Certifications
Quality System
Download QMSQMS (ISO, GMP)
Download CertificateQuality Advantages
View ReportQuality Control Process
Download ProcessTechnical Resources & Downloads
Frequently Asked Questions (FAQ)
Q1: What is the required sample input and preparation?
Start from freshly harvested 10^6 cells with viability ≥80% for extraction. Rinse cells with PBS, lyse with Cell Lysis Buffer Working Solution, incubate on ice 30 minutes, centrifuge and collect supernatant as extract. For high-activity samples consider 10-fold dilution or extracting from 10^5 cells.
Q2: What is the assay dynamic range and detection limit?
The standard series covers 0.4 to 400,000 TPG Units (ST6–ST0). The lowest standard provided corresponds to 0.4 TPG Units (0.0002 amol/μL TSR8). Confirm instrument-specific LOD/LOQ during method validation.
Q3: How should samples and reagents be stored?
Store kit components at –20 °C (protect from light where indicated). Cell pellets can be stored at –70 °C up to one year; prepared extracts should be aliquoted and kept at –70 °C (no more than 6 months) to avoid freeze–thaw.
Q4: What controls are required in each run?
Include No Template Control (NTC), Heat-treated negative quality control (HT sample), Positive quality control (telomerase positive cell extract) and a TSR8 standard curve (at least five concentrations). Internal control (CY5) monitors inhibition.
Research Use Only
✔
For Research Use Only (RUO) – This product is intended for laboratory research purposes only and not for clinical or regulatory diagnostic use.
✔
Not intended for use in USDA or FDA regulated diagnostic testing or official compliance testing.
When can I expect my order to ship?
Most orders are filled and shipped within 2-3 business days from the time they are received.
Our standard shipping usually take 2-5 days.
We also provide express shippping for time-sensitive deliveries.
Email contact@biofargo.com if you have any requirements.
