| Field | Specification |
|---|---|
| Mfr No | |
| Alternative Names | TPPP/p25; TPPP1; p24; p25; p25alpha; brain specific protein p25 alpha|glycogen synthase kinase 3 (GSK3) inhibitor p24|tubulin polymerization-promoting protein |
| Product Type | |
| Sensitivity | |
| UniProt # |
Scientific Background
Seki et al. (1999) cloned p25 from a neuroblastoma cDNA library. The deduced 219-amino acid protein has a calculated molecular mass of about 24 kD. Human and bovine p25 share 90% amino acid identity. In contrast to the brain-specific expression of bovine p25, RT-PCR detected human p25 in all tissues examined.Martin et al. (2002) found that p24 associated with Gsk3 in microtubules from adult rat brain. p24 bound Gsk3 and inhibited its kinase activity at low magnesium concentrations. p24 was a poor substrate for Gsk3, but it could be phosphorylated by other protein kinases. Bovine Tppp inhibited.
Assay Principle
This assay employs a two-site sandwich ELISA to quantitate TPPP in samples. An antibody specific for TPPP has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyTPPP present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for TPPP 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 TPPP bound in the initial step. The color development is stopped and the intensity of the color is measured.
Performance Specifications
| Sensitivity | 0.128 ng/mL |
|---|---|
| Detection Range | 0.312-20 ng/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 0.128 ng/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 TPPP 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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