Key Features & Benefits
- Enhanced Sensitivity with Superior Detection: Utilizes >95% pure biotinylated recombinant monoclonal antibodies paired with streptavidin-HRP conjugate (activity ≥500 U/mg) for amplified signal and a low detection limit of <0.01 ng/mL.
- High Specificity for Active Metabolite: Accurately measures 1α,25-(OH)2D3 with minimal cross-reactivity to other Vitamin D metabolites (e.g., 25-OH-D2/D3), ensuring reliable data.
- Optimized & Streamlined Protocol: A user-friendly one-step assay reduces hands-on time and simplifies the workflow for efficient processing of multiple samples.
- Exceptional Value Proposition: Obtain premium performance for less. Our Human 1α,25-(OH)2D3 ELISA Kit for sale comes with wholesale price starting under $100.
- Comprehensive Research Support: We offer custom ELISA kit development for unique project needs and a complimentary sample testing service to verify assay performance with your specific samples.
Enhanced Assay Principle: Biotin-Streptavidin Amplified ELISA
This kit utilizes a sensitive one-step sandwich enzyme-linked immunosorbent assay (ELISA) with signal amplification:
- Capture: A specific antibody for 1α,25-(OH)2D3 is pre-coated onto the microplate wells.
- Formation of Sandwich Complex: Samples or standards are incubated simultaneously with a biotinylated detection antibody. The target 1α,25-(OH)2D3 is bound by both antibodies.
- Signal Amplification: After washing, a Streptavidin-Horseradish Peroxidase (SA-HRP) conjugate is added, which binds with high affinity to the biotin on the detection antibody, creating an amplified detection complex.
- Color Development & Measurement: Following another wash, TMB substrate is added. The HRP catalyzes TMB’s conversion to a blue product, which turns yellow upon acid addition. The absorbance at 450 nm is measured. The signal intensity is directly proportional to the concentration of 1α,25-(OH)2D3 in the sample.
Detailed Kit Specifications & Contents
Performance Characteristics
| Parameter | Specification |
|---|---|
| Accuracy (Linearity) | Correlation coefficient R ≥ 0.9900 |
| Sensitivity | < 0.01 ng/mL |
| Detection Range | 0.1 – 2 ng/mL |
| Specificity | No significant cross-reactivity with analogous molecules |
| Precision (Repeatability) | Intra- and Inter-assay CV < 15% |
| Storage | 2-8°C, protect from light and moisture |
| Shelf Life | 6 months |
Standard Curve Concentrations
S0 to S5: 0, 0.125, 0.25, 0.5, 1, 2 ng/mL
Complete Kit Components (96T / 48T)
- Pre-coated Microplate (8 strips / 4 strips)
- Standards (0.3mL x 6 tubes)
- Sample Diluent (6mL / 3mL)
- Biotinylated Detection Antibody (10mL / 5mL)
- Streptavidin-HRP Conjugate (10mL / 5mL)
- 20x Wash Buffer (25mL / 15mL)
- Substrate A & B (6mL each / 3mL each)
- Stop Solution (6mL / 3mL)
- Plate Sealers, Comprehensive Manual, Storage Bag
Sample Handling & Preparation Guidelines
- Serum/Plasma: Collect using sterile, pyrogen-free tubes. Centrifuge at 3000×g for 10-30 minutes to obtain a clear supernatant free of cells or particles.
- Cell Culture Supernatant & Tissue Homogenate: Clarify by centrifugation at 3000×g for 10 minutes.
- Storage: For long-term storage, aliquot samples and freeze at -20°C. Avoid repeated freeze-thaw cycles. Thaw completely at room temperature and mix well before assay.
Essential Workflow & Notes
- Preparation: Equilibrate all kit components to room temperature for 20 minutes. Dilute the 20x Wash Buffer 1:20 with distilled water.
- Sample & Standard Addition: Add 50µL of standards or pre-diluted samples (10µL sample + 40µL diluent) to appropriate wells. The S0 standard serves as the blank/zero control.
- Incubation with Detection Reagents: Add 100µL of the prepared detection antibody/SA-HRP mixture to each well (except blank). Incubate at 37°C for 60 minutes.
- Washing: Wash the plate 5 times thoroughly using the prepared wash buffer.
- Substrate Reaction: Add 50µL each of Substrate A and B. Incubate at 37°C for 15 minutes in the dark.
- Stop & Read: Add 50µL Stop Solution. Measure the absorbance at 450 nm within 15 minutes.
Note: Final sample concentration = Calculated concentration from curve × dilution factor (5).
Materials Required But Not Supplied
- Microplate Reader (450 nm filter)
- Precision Pipettes and Disposable Tips
- 37°C Incubator or Water Bath
- Deionized or Distilled Water
Primary Research Applications
- Bone & Mineral Metabolism Research: Investigate the role of active Vitamin D in calcium regulation, osteoporosis, and renal osteodystrophy.
- Immunology & Autoimmunity: Study the immunomodulatory effects of 1α,25-(OH)2D3 in diseases like multiple sclerosis, rheumatoid arthritis, and immune cell differentiation.
- Cancer Research: Explore correlations between Vitamin D status, receptor expression, and outcomes in various cancers.
- Endocrine & Nutritional Studies: Assess Vitamin D sufficiency, metabolism, and its impact on overall health.
Ordering, Customization & Value-Added Services
Purchase the Standard Kit: Buy the Human 1α,25-(OH)2D3 ELISA Kit online from Yanda Bio. Contact us for a specific quote, with bulk order pricing maintained under $100.
Custom ELISA Development Service: Require a modified assay range, different sample type validation, or unique conjugate? Our R&D team can develop a tailored solution.
Free Sample Testing Service: Validate the kit’s applicability for your project risk-free. Send us your samples; we will run the assay and provide a complete data report.
Choose Yanda Bio for reliable tools and expert support. Contact us today to order, request a custom quote, or schedule your complimentary sample analysis.
Frequently Asked Questions (FAQ)
Q: What is the advantage of the biotin-streptavidin detection system?
A: This system provides significant signal amplification. Multiple biotin molecules on the detection antibody bind multiple streptavidin-HRP complexes, greatly enhancing sensitivity and allowing detection of very low analyte levels (<0.01 ng/mL).
Q: Can this kit distinguish between 1α,25-(OH)2D2 and D3?
A: This kit is specifically designed for the D3 form (cholecalciferol-derived). Please check the cross-reactivity data in the manual or contact us for specifics regarding D2.
Q: How do you offer such competitive pricing?
A: Our optimized manufacturing and direct-to-researcher sales model eliminate intermediary costs, allowing us to offer high-performance Human 1α,25-(OH)2D3 ELISA Kits for sale at wholesale prices under $100.
Q: Is this kit suitable for clinical diagnosis?
A: No. This product is for Research Use Only (RUO). It is not intended for use in diagnostic procedures.
Disclaimer
- This kit is designed for research purposes only. It is not validated or approved for use in clinical, diagnostic, or therapeutic procedures involving humans.
- Users must adhere strictly to the provided instructions. The manufacturer is not liable for consequences arising from the misuse or modification of the prescribed protocol.
