| Field | Specification |
|---|---|
| Mfr No | |
| Alternative Names | MGC11268; pseudouridine synthase 1|tRNA uridine isomerase I |
| Product Type | |
| Sensitivity | |
| UniProt # |
Scientific Background
PUS1 converts uridine into pseudouridine after the nucleotide has been incorporated into RNA. Pseudouridine may have a functional role in tRNAs and may assist in the peptidyl transfer reaction of rRNAs PUS1-1 encodes a protein with an N-terminal mitochondrial leader peptide. Western blot analysis detected full-length PUS1-1 at an apparent molecular mass of 47 kD, and the mature protein, lacking the mitochondrial localization signal, was about 37 kD. The PUS1-2 and PUS1-3 transcripts differ in the 5-prime untranslated region but encode the same protein that begins with an internal methionine (m.
Assay Principle
This assay employs a two-site sandwich ELISA to quantitate PUS1 in samples. An antibody specific for PUS1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyPUS1 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for PUS1 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 PUS1 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Performance Specifications
| Sensitivity | 13.6 pg/mL |
|---|---|
| Detection Range | 31.25-2000 pg/mL |
| Total Assay Time | 3.5–5 hours |
| Compatible Sample Types | serum, plasma, cell culture supernatant, tissue homogenate |
| Species Reactivity | Rat |
| 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 13.6 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 PUS1 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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