Target Identity Clarification
This kit measures 2′,3′-cyclic nucleotide 3′-phosphodiesterase (CNPase), a myelin-associated enzyme abundant in the central nervous system. It does not detect C-type natriuretic peptide (CNP). Please verify that CNPase is your intended target before ordering.
Key Features
- Authentic CNPase Detection: Specifically quantifies human 2′,3′-cyclic nucleotide 3′-phosphodiesterase, a key structural protein of myelin.
- Optimized for Intracellular Protein: Includes guidance for tissue and cell lysis using protease inhibitor-supplemented RIPA buffer to efficiently release the cytoplasmic target.
- High Sensitivity for Low-Abundance Samples: Minimum detectable concentration < 1.0 pg/mL; quantitative range 1.0 – 80 pg/mL, covering the low endogenous levels found in biological fluids.
- Validated Sample Types: Human serum, EDTA/citrate/heparin plasma, cell culture supernatant, and tissue homogenates. Tissue or cell lysates are strongly recommended due to the intracellular nature of CNPase.
- Built-in 5× Sample Dilution: 10 µL sample + 40 µL Sample Diluent in-well; multiply the interpolated result by 5 to obtain the actual concentration.
- Recombinant Standard: Produced in a eukaryotic expression system; ready-to-use liquid format (0, 5, 10, 20, 40, 80 pg/mL).
- One-Step Sandwich ELISA: Simultaneous incubation of sample and HRP-detection antibody; total assay time approximately 2 hours.
- Excellent Precision: Intra- and inter-assay CV < 15%; standard curve linearity R ≥ 0.9900.
- Export-Ready: Neutral white box (16 × 12.5 × 9.5 cm), cold-chain transport (2–8°C), 6-month shelf life. MSDS and Certificate of Composition available upon request.
Product Description
2′,3′-Cyclic nucleotide 3′-phosphodiesterase (CNPase) constitutes approximately 4% of total myelin protein in the central nervous system. It catalyzes the hydrolysis of 2′,3′-cyclic nucleotides to 2′-nucleotides and is widely used as a marker of oligodendrocyte differentiation, myelination, and demyelinating disorders. In human research, CNPase quantitation supports studies in multiple sclerosis, brain injury, and neural development.
The Yanda-HUMAN-CNPase ELISA Kit employs a one-step double-antibody sandwich ELISA. A capture antibody specific for human CNPase is pre-coated on the microplate. Samples and standards are co-incubated with an HRP-labeled detection antibody. After washing, TMB substrate generates a colorimetric signal proportional to CNPase concentration, measured at 450 nm.
Sample Selection Note: Because CNPase is an intracellular protein predominantly expressed in myelin, its concentration in serum or plasma is naturally very low and may approach or fall below the assay’s detection limit. For best results, use cell lysates or tissue homogenates (especially brain tissue) as your primary sample type.
Sample Collection & Preparation
Recommended Sample Types
| Sample Type | Preparation |
|---|---|
| Tissue Homogenate (e.g., brain) | Strongly recommended. Homogenize in ice-cold RIPA buffer containing fresh protease inhibitors. Incubate on ice 30 min, centrifuge at 12,000 × g for 15 min at 4°C, collect supernatant. |
| Cell Lysate | Wash cells with cold PBS, lyse with RIPA buffer + protease inhibitors, incubate on ice 30 min, centrifuge at 12,000 × g for 15 min at 4°C, collect supernatant. |
| Serum / Plasma | Collect as per standard protocols. Note: free CNPase in blood is extremely low; a negative or near-blank result is expected in healthy donors. |
| Cell Culture Supernatant | Centrifuge at 3000 rpm for 10 min to remove debris. Secreted CNPase levels are typically very low; lysis of the cell pellet is recommended for meaningful quantitation. |
General Guidelines
- Protease inhibitors: Mandatory for all tissue/cell lysates to prevent degradation.
- Avoid freeze-thaw: Aliquot and store at -20°C. Do not repeatedly freeze-thaw.
- NaN3 prohibition: Sodium azide inhibits HRP and must not be present in samples.
Critical Dilution Factor
The assay performs a 5-fold dilution in the well (10 µL sample + 40 µL Sample Diluent). The value interpolated from the standard curve is the concentration in the diluted sample. Multiply this value by 5 to obtain the concentration in the original specimen.
The S0 standard (0 pg/mL) serves as the blank/negative control.
Pre-dilution note: For tissue/cell lysates with high total protein content, a pilot experiment with several dilution factors is recommended to ensure readings fall within the 1.0 – 80 pg/mL range. If values exceed 80 pg/mL after the 5× correction, apply additional pre-dilution and multiply by the extra factor.
Technical Specifications
| Parameter | Specification |
|---|---|
| Standard Concentrations (S0–S5) | 0, 5, 10, 20, 40, 80 pg/mL |
| Quantitative Range | 1.0 – 80 pg/mL |
| Analytical Sensitivity | < 1.0 pg/mL |
| Accuracy (Linearity) | Standard curve R ≥ 0.9900 |
| Precision | Intra-assay CV < 15%; Inter-assay CV < 15% |
| Specificity | Highly specific for human CNPase. No cross-reactivity with other soluble structural analogues, including C-type natriuretic peptide (CNP). |
| Sample Dilution Factor | 5× (in-well); additional pre-dilution may be needed |
| Shelf Life | 6 months |
| Storage & Transport | 2–8°C, protected from light and moisture; cold-chain shipping |
Representative Standard Curve
Detailed validation data are available in the product instruction manual. Please contact technical support for further information.
Kit Components
96-Well Format
| Component | Quantity | Notes |
|---|---|---|
| Microelisa Stripplate | 12 strips × 8 wells | Pre-coated with anti-human CNPase antibody |
| Standard (0.3 mL/vial) | 6 vials | Ready-to-use liquid, 0–80 pg/mL |
| Sample Diluent | 6 mL | – |
| HRP-Conjugated Detection Antibody | 10 mL | Ready-to-use |
| 20× Wash Buffer | 25 mL | Dilute to 1× before use |
| Substrate A | 6 mL | – |
| Substrate B | 6 mL | – |
| Stop Solution | 6 mL | – |
| Plate Sealers | 2 sheets | – |
| Instruction Manual | 1 copy | – |
| Self-sealing Bag | 1 piece | For unused strip storage |
48-Well Format
| Component | Quantity | Notes |
|---|---|---|
| Microelisa Stripplate | 12 strips × 4 wells | Pre-coated, removable |
| Standard (0.3 mL/vial) | 6 vials | Ready-to-use liquid |
| Sample Diluent | 3 mL | – |
| HRP-Conjugated Detection Antibody | 5 mL | Ready-to-use |
| 20× Wash Buffer | 15 mL | Dilute to 1× |
| Substrate A | 3 mL | – |
| Substrate B | 3 mL | – |
| Stop Solution | 3 mL | – |
| Plate Sealers | 2 sheets | – |
| Instruction Manual | 1 copy | – |
| Self-sealing Bag | 1 piece | – |
Crystallization of 20× Wash Buffer upon cold storage is normal; warm in a water bath to dissolve completely before dilution.
Assay Procedure Summary
- Equilibrate: Bring the sealed foil pouch to room temperature for 20 min. Remove required strips; reseal unused strips and store at 4°C.
- Add Standards & Samples:
- Standard wells: 50 µL of each standard.
- Sample wells: 10 µL of sample + 40 µL Sample Diluent.
- Blank well(s): leave empty.
- Add Detection Antibody: Add 100 µL HRP-conjugated detection antibody to all wells except blank. Seal and incubate at 37°C for 60 minutes.
- Wash: Aspirate, blot dry. Fill wells with 1× Wash Buffer, soak 1 min, discard. Repeat 5 times (automated: 350 µL/well).
- Substrate: Add 50 µL each of Substrate A and Substrate B. Incubate in the dark at 37°C for 15 minutes.
- Stop & Read: Add 50 µL Stop Solution. Read absorbance at 450 nm within 15 min.
Total assay time: approximately 2 hours.
Materials Required but Not Supplied:
- Microplate reader (450 nm)
- Adjustable pipettes (0.5–1000 µL)
- 37°C incubator or water bath
- Distilled/deionized water
- Absorbent paper, wash bottle or automated washer
- RIPA buffer, protease inhibitors (for tissue/cell lysis)
Throughput
| Format | Max. Samples (Single Wells) |
|---|---|
| 48-well | 39–40 |
| 96-well | 85–88 |
Includes 6 standard wells and 1–2 blank wells. For duplicates, divide the sample number accordingly.
Frequently Asked Questions
Q: Is this kit for C-type natriuretic peptide (CNP)?
A: No. This kit detects 2′,3′-cyclic nucleotide 3′-phosphodiesterase (CNPase), a myelin enzyme, not the natriuretic peptide CNP. Please carefully check the target before purchase.
Q: Which sample type should I use?
A: Because CNPase is an intracellular protein, tissue homogenates (especially brain) or cell lysates are strongly recommended. CNPase levels in serum or plasma are extremely low and may be undetectable.
Q: How do I prepare tissue or cell lysates for CNPase detection?
A: Use RIPA buffer containing fresh protease inhibitors. Homogenize or lyse on ice, incubate for 30 min, and centrifuge at 12,000 × g for 15 min at 4°C. Use the supernatant for the assay.
Q: Does this kit cross-react with CNP or other phosphodiesterases?
A: No. The kit is highly specific for CNPase and does not cross-react with C-type natriuretic peptide (CNP) or other soluble structural analogues.
Q: What if my serum sample gives a negative result?
A: This is expected for many healthy individuals due to the low circulating concentration of this intracellular protein. We recommend using tissue or cell lysates for meaningful CNPase quantitation.
Q: Can I use this kit for animal samples?
A: The kit has been validated only for human samples. Cross-reactivity with other species has not been established.
Q: Do you provide MSDS and composition documents for export?
A: Yes. These documents are available upon request and can be included with the shipment.
Ordering & Contact Information
| Item | Details |
|---|---|
| Catalog Number | Yanda-HUMAN-CNPase |
| Formats | 48-well / 96-well |
| Package Dimensions | 16 cm × 12.5 cm × 9.5 cm (neutral white box) |
| Shipping | Cold-chain (2–8°C), protected from light and moisture |
| Storage | 2–8°C; do not freeze |
| Shelf Life | 6 months |
| Intended Use | For research use only, not for diagnostic procedures |
Contact us
- WhatsApp: +86 198 3186 0669
- Email: songhawyble@gmail.com
For pricing, bulk orders, or export documentation, please contact us directly.
Technical Documents & Support
- MSDS & Certificate of Composition – Available upon request.
- Technical Support – For protocol assistance or validation data, reach out via WhatsApp or email.
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Disclaimer
This product is for laboratory research use only and is not approved for clinical diagnosis, treatment, or prognosis. The manufacturer assumes no responsibility for consequences arising from improper use, protocol deviations, or unauthorized applications. Users are responsible for validating assay performance in their own sample matrices and experimental conditions.
