| Field | Specification |
|---|---|
| Mfr No | |
| Alternative Names | 53BP1; FLJ41424; MGC138366; p202; tumor protein 53-binding protein; 1|tumor protein p53-binding protein; 1 |
| Product Type | |
| Sensitivity | |
| UniProt # |
Scientific Background
TP53BP1 has a predicted molecular mass of 217 kD and shows homology in its C-terminal 247 amino acids to the C terminus of the tumor suppressor protein BRCA1. TP53BP1 was present in both the cytoplasm and nucleus in some cells and only in the nucleus in others. Furthermore, there were 2 nuclear staining patterns, one homogeneous staining and the other dot staining.TP53BP1 was required for p53 accumulation, G2/M checkpoint arrest, and the intra-S-phase checkpoint in response to ionizing radiation. TP53BP1 played a partially redundant role in phosphorylation of the downstream checkpoint effector.
Assay Principle
This assay employs a two-site sandwich ELISA to quantitate TP53BP1 in samples. An antibody specific for TP53BP1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyTP53BP1 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for TP53BP1 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 TP53BP1 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Performance Specifications
| Sensitivity | 0.138 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 0.138 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 TP53BP1 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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