| Field | Specification |
|---|---|
| Mfr No | |
| Alternative Names | RP11-68I18.9; |
| Product Type | |
| Sensitivity | |
| UniProt # |
Scientific Background
Phosphatidylinositol-4-phosphate 5-kinases synthesize phosphatidylinositol 4,5-bisphosphate by phosphorylating phosphatidylinositol 4-phosphate. The deduced 549-amino acid protein has the conserved kinase homology domain of PIP5K family members. Within this domain, PIP5K1A shows 83% and 35% amino acid identity with PIP5K1B and PIP5K2A , respectively. Overall, the PIP5K1A and PIP5K1B proteins are 64% identical. Recombinant PIP5K1A expressed in bacteria had a molecular mass of approximately 66.3 kD. The authors isolated additional PIP5K1A cDNAs which they suggested represent splicing isoforms. N.
Assay Principle
This assay employs a two-site sandwich ELISA to quantitate PIP5K1A in samples. An antibody specific for PIP5K1A has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyPIP5K1A present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for PIP5K1A 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 PIP5K1A bound in the initial step. The color development is stopped and the intensity of the color is measured.
Performance Specifications
| Sensitivity | 34.8 pg/mL |
|---|---|
| Detection Range | 62.5-4000 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 34.8 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 PIP5K1A 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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