| Field | Specification |
|---|---|
| Mfr No | |
| Accession Number | |
| Alternative Names | TPPP; TPPP/p25; TPPP1; p24; p25; p25alpha; brain specific protein p25 alpha; glycogen synthase kinase 3 (GSK3) inhibitor p24; tubulin polymerization-promoting protein |
| Assay Time | |
| Assay Type | |
| Detection Method | |
| Gene ID | |
| Product Type | |
| Reactivity | |
| Sample Type(s) | Cell culture supernatants, Serum, Plasma, Other biological fluids |
| Shipping | |
| Storage |
Features & Benefits
Human Tubulin polymerization-promoting protein (TPPP) ELISA Kit has high sensitivity and excellent specificity for detection of Human TPPP. No significant cross-reactivity or interference between Human TPPP and analogues was observed.
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 mitotic spindle assembly and nuclear envelope breakdown following injection into Drosophila embryos. It did not affect other cellular events.
This Tubulin polymerization-promoting protein (TPPP) ELISA kit is validated for use with Cell culture supernatants, Serum, Plasma, Other biological fluids. Samples should be collected, processed, and stored correctly to preserve analyte integrity — avoid repeated freeze-thaw cycles and centrifuge to remove particulates before use. Dilute samples exceeding the kit's detection range using the supplied assay diluent. Hemolytic, icteric, or lipemic samples may affect assay performance and should be tested with caution.
This is a sandwich ELISA kit employing an HRP (horseradish peroxidase)-conjugated secondary antibody paired with a TMB (3,3′,5,5′-tetramethylbenzidine) colorimetric substrate. In the sandwich format, the target analyte Tubulin polymerization-promoting protein (TPPP) captured on the microplate surface is detected by the conjugated antibody, generating a colorimetric signal proportional to analyte concentration. The reaction is stopped and absorbance measured at 450 nm on a standard microplate reader.
The complete protocol, from sample addition to final plate reading, requires approximately 3–5 hours. This includes two incubation periods (analyte binding and detection antibody steps), intermediate wash cycles to remove unbound material, 15–30 minutes of TMB substrate development, and final stop-solution addition before absorbance reading. Exact timing will vary with experience level and the number of samples processed in parallel.
Required equipment: (1) a microplate spectrophotometer capable of reading absorbance at 450 nm (reference wavelength 540–570 nm recommended for background correction); (2) precision single-channel or multichannel pipettes; (3) a plate washer or multichannel aspirator; (4) a microcentrifuge for sample clarification; and (5) a 37°C incubator or stable room-temperature environment. No fluorescence or luminescence reader is required — standard colorimetric plate readers are fully compatible with this kit.
This ELISA kit is formally validated for Human. Cross-reactivity with species not listed in the specification has not been independently characterized. Variability in protein sequence homology across species means that performance in unlisted species cannot be guaranteed without additional validation. For cross-species detection requirements or non-standard sample matrices, please contact BioHippo support or refer to the manufacturer's technical team for guidance.
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