| Field | Specification |
|---|---|
| Mfr No | |
| Alternative Names | AP4; RIP60; ZNF464; Zfp464; H_DJ0584D14.12|replication initiation region protein (60kD)|zinc finger protein 464 (RIP60)|zinc finger protein AP4|zinc finger proten AP4 |
| Product Type | |
| Sensitivity | |
| UniProt # |
Scientific Background
Replication initiator 1 is a protein encoded by the REPIN1 gene. Homodimers of RIP60 (replication initiation-region protein 60 kDA) purified from nuclear extract bind two ATT-rich sites in oribeta and foster the formation of a twisted 720 bp DNA loop in vitro. RIP60 contains 15 C(2)H(2)zinc finger (ZF) DNA binding motifs organized in three clusters, termed hand Z1 (ZFs 1-5), hand Z2 (ZFs 6-8) and hand Z3 (ZFs 9-15). A proline-rich region is located between hands Z2 and Z3. Gel mobility shift and DNase I footprinting experiments show hands Z1 and Z2 independently bind the oribeta RIP60 sites sp.
Assay Principle
This assay employs a two-site sandwich ELISA to quantitate REPIN1 in samples. An antibody specific for REPIN1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyREPIN1 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for REPIN1 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 REPIN1 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Performance Specifications
| Sensitivity | 0.129 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 0.129 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 REPIN1 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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