| Field | Specification |
|---|---|
| Applications | |
| Assay type | |
| Sample type(s) | Serum | Plasma | Other Biological Fluid Samples |
| Sensitivity | |
| Detection range | |
| Detection method | |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Product Overview
The Anti-Sacituzumab Neutralizing Antibody ELISA Kit is a competitive ELISA kit for quantitative measurement of neutralizing anti-sacituzumab antibodies in serum, plasma and other biological fluid samples. The 96-well microplate is pre-coated, and the colorimetric signal is read at 450 nm.
In this competitive format, neutralizing anti-sacituzumab antibodies in the sample block the binding of sacituzumab to its target in the coated wells, so the optical density falls as the neutralizing antibody level rises. Results are calculated from the standard curve run on the same plate.
Values are expressed relative to the antibody standard supplied with the kit.
Specifications
| Assay Type | Competitive (Quantitative) |
|---|---|
| Detection Method | Colorimetric (OD 450 nm) |
| Sample Type | Serum, Plasma, Other Biological Fluid Samples |
| Sensitivity | 16.06 ng/mL |
| Detection Range | 31.25 - 2,000 ng/mL |
| Recovery | 80-120% |
| Precision | Intra-assay CV <15%; Inter-assay CV <15% |
| Kit Size | 96 T |
| Shipping | 2-8 °C |
Kit Components
| Component | Quantity | Storage |
|---|---|---|
| Pre-coated Microplate | 1 plate | -20 °C |
| Standard | 2 bottles | -20 °C |
| Detection A | 1 vial | -20 °C |
| Detection B | 1 vial | -20 °C |
| Standard Diluent | 1 bottle | 4 °C |
| Assay Diluent | 1 bottle | 4 °C |
| 20× Wash Buffer | 1 bottle | 4 °C |
| Color Reagent | 1 bottle | 4 °C |
| Stop Solution | 1 bottle | 4 °C |
| Plate Sealers | 4 strips | Room temperature |
Assay Procedure (Summary)
Bring all reagents to room temperature and prepare working solutions and the standard series before starting.
- Add 100 µL standard or sample premixed with Detection A working solution (50 µL + 50 µL); incubate 1 h at 37 °C.
- Wash 3 times with 300 µL wash buffer per well.
- Add 100 µL Detection B working solution; incubate 30 min at 37 °C.
- Wash 5 times with 300 µL wash buffer per well.
- Add 100 µL Color Reagent; incubate 15 min at 37 °C, protected from light.
- Add 50 µL stop solution; the color turns from blue to yellow.
- Read the OD at 450 nm within 10 min (wavelength correction at 630 nm or 620 nm).
Total incubation time is about 1 h 45 min, plus washing steps. Required but not supplied: a microplate reader for 450 nm (correction at 630 nm or 620 nm), pipettes and tips, deionized or distilled water, a 500 mL graduated cylinder, a squirt bottle, manifold dispenser or plate washer, and tubes for standard dilutions.
Sample Preparation
- Serum: use a serum separator tube, let the sample clot for 30 minutes at room temperature, and centrifuge for 15 minutes at 1000 × g.
- Plasma: collect with EDTA, heparin or citrate as the anticoagulant and centrifuge for 15 minutes at 1000 × g within 30 minutes of collection.
- Assay samples promptly or store aliquots at -20 °C or below; avoid repeated freeze-thaw cycles.
- Serum and plasma samples: a 10-fold starting dilution in Standard Diluent is suggested; adjust it so readings fall within the standard curve.
Storage & Handling
For Research Use Only. Not for use in diagnostic procedures.
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