| Field | Specification |
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Scientific Background
Thymidine kinase (EC 2.7.1.21) catalyzes the phosphorylation of thymidine to deoxythymidine monophosphate. Genetic variation in soluble thymidine kinase has not been identified in man. Weiss and Green (1967) found that fusion of mouse cells lacking this enzyme with normal human cells could be achieved, that progressive loss of human chromosomes from the hybrid occurred with passage of time, and that at a stage when only one human chromosome remained the cell still had the capacity to synthesize thymidine kinase. Two forms of Thymidine kinase 1, soluble have been identified in animal cells, one.
Assay Principle
This assay employs a two-site sandwich ELISA to quantitate TK2 in samples. An antibody specific for TK2 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyTK2 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for TK2 is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of TK2 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Performance Specifications
| Sensitivity | 6.3 pg/mL |
|---|---|
| Detection Range | 15.6-1000 pg/mL |
| Total Assay Time | 3.5–5 hours |
| Compatible Sample Types | serum, plasma, cell culture supernatant, tissue homogenate |
| Species Reactivity | Human |
| Detection Method | Colorimetric (TMB/HRP) |
| Storage | 4°C (short-term); -20°C (long-term) |
✓ Research-Grade Validation
Safety & Regulatory
Handle reagents in accordance with institutional biosafety guidelines. Refer to the Safety Data Sheet (SDS) for complete hazard and handling information. Contains components that may require special disposal procedures per local regulations.
This kit is validated for use with serum, plasma, cell culture supernatant, tissue homogenate. For unlisted matrices (e.g., tissue lysate, urine), perform a spike-and-recovery experiment to confirm assay performance before generating reportable data. Sample dilution in the kit's provided diluent is recommended to minimize matrix interference.
The minimum detectable concentration (sensitivity) of this kit is 6.3 pg/mL. Values below this threshold should be reported as below the limit of detection (<LOD) and should not be extrapolated from the standard curve.
The total assay time from sample addition to absorbance reading is approximately 3.5–5 hours, including all incubation, wash, and substrate steps. Hands-on time is typically 1–2 hours; most steps involve passive plate incubation. Plan the assay as a single uninterrupted session for best results.
Standard components of this Sandwich ELISA Kit typically include: pre-coated microplate (96-well strip format), lyophilized or liquid recombinant TK2 standard, detection antibody, streptavidin-HRP conjugate, TMB substrate, stop solution, wash buffer concentrate, and sample/standard diluent. Refer to the kit insert or datasheet for the exact component list and storage requirements.
This kit uses colorimetric (TMB/HRP) detection and requires a standard microplate absorbance reader capable of measuring at 450 nm. A reference wavelength of 570 nm or 630 nm is recommended to reduce background. No specialized fluorescence or luminescence reader is needed. Ensure the instrument is calibrated and the plate is clean and free of condensation before reading.
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