Target Distinction
This kit specifically measures rabbit Type I collagen (Col-I) , the most abundant fibrillar collagen in skin, bone, tendon, and ligaments. It is not a pan-collagen assay and does not detect total collagen or other collagen types (II, III, IV, etc.). If you require a human total collagen kit, please refer to our separate product (Yanda-HUMAN-Col).
Key Features
- Specific for Native Rabbit Type I Collagen: Only identify the unique antigenic epitopes specific to Rabbit Type I Collagen.Detects the natural triple-helical conformation of Col-I. No significant cross-reactivity with other collagen types or ECM proteins.
- Optimized for Rabbit Models: Validated exclusively for rabbit samples (serum, plasma, tissue homogenates, cell lysates, and cell culture supernatant).
- Practical Pre-Dilution Guide: Serum/plasma typically requires 1:2–1:8 dilution; tissue homogenates from fibrotic models may need 1:20 or higher. Detailed pilot testing recommendations provided.
- Tissue Extraction Flexibility: Standard PBS homogenization is suitable for soluble collagen. For highly cross-linked mature collagen fibers, pepsin digestion may be employed based on your extraction efficiency needs.
- High Sensitivity & Defined Range: Quantitative range 1.0–40 ng/mL, sensitivity <1.0 ng/mL. Standard curve: 0, 2.5, 5, 10, 20, 40 ng/mL.
- Built-in 5× In-Well Dilution: 10 µL sample + 40 µL Sample Diluent; multiply the interpolated result by 5 to obtain the actual concentration.
- Ready-to-Use Liquid Standards: Recombinant protein standards produced in a eukaryotic expression system; no reconstitution needed.
- One-Step Sandwich ELISA: Co-incubation of sample and HRP-detection antibody; total assay time ~2 hours.
- Reliable 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 shipping (2–8°C), 6-month shelf life. MSDS available upon request.
Product Description
Type I collagen is the primary structural protein of the extracellular matrix, constituting over 90% of organic bone matrix and providing tensile strength to tendons, ligaments, and dermis. In rabbit models, Col-I is a critical biomarker for studying wound healing, bone regeneration, tissue fibrosis (liver, lung, kidney), and scleroderma. Its massive accumulation during fibrogenesis makes accurate quantitation essential for evaluating anti-fibrotic therapies.
The Yanda-Rabbit-Col-I ELISA Kit employs a one-step double-antibody sandwich ELISA. A capture antibody specific for native rabbit Col-I is pre-coated on the microplate. Samples and standards are co-incubated with an HRP-labeled detection antibody. After washing, TMB substrate produces a colorimetric signal directly proportional to the Col-I concentration, measured at 450 nm.
Important: This kit detects native Type I collagen, not recombinant or denatured fragments. It is validated for rabbit samples only and does not cross-react with other species.
Sample Collection & Preparation
Validated Sample Types
| Sample Type | Collection & Processing |
|---|---|
| Serum | Collect in pyrogen/endotoxin-free tubes. Allow to clot, centrifuge at 3000 rpm for 10 min. Separate serum immediately. |
| Plasma | Use EDTA, citrate, or heparin. Centrifuge at 3000 rpm for 30 min; collect supernatant. |
| Cell Culture Supernatant | Centrifuge at 3000 rpm for 10 min to remove debris. |
| Cell Lysate | Wash cells with cold PBS, lyse with appropriate buffer, centrifuge at 3000 rpm for 10 min at 4°C. Collect supernatant. |
| Tissue Homogenate | See detailed protocol below. |
Tissue Homogenate Preparation
Standard Method:
- Rinse fresh tissue with ice-cold PBS (0.01 M, pH 7.4) to remove residual blood.
- Mince tissue, add PBS at a ratio of 1:9 (w/v), and homogenize thoroughly on ice using a glass homogenizer or tissue grinder.
- Centrifuge at 3000 rpm for 10 min at 4°C. Collect the supernatant for testing.
For Highly Cross-Linked Mature Collagen (e.g., Fibrotic Tissues):
Standard PBS homogenization may yield low extraction efficiency due to extensive covalent cross-linking. If your research targets insoluble mature collagen fibers, consider adding pepsin digestion (e.g., 0.5 M acetic acid containing pepsin at 1 mg/mL, incubated overnight at 4°C with gentle agitation). After digestion, neutralize the pH, centrifuge, and use the supernatant for the assay. We recommend running a pilot comparison between PBS-only and pepsin-digested extracts to determine which protocol better reflects your target analyte pool.
Storage
If not tested immediately, aliquot samples and store at -20°C. Avoid repeated freeze-thaw cycles. Thaw completely at room temperature and mix gently before use.
Pre-Dilution Recommendations
Because Col-I concentrations vary enormously across sample types and disease states, pre-experimental gradient dilution is essential for optimal results.
| Sample Type | Suggested Pre-Dilution Range | Remarks |
|---|---|---|
| Normal serum / plasma | 1:2, 1:4, 1:8 | Baseline levels typically in the low ng/mL range |
| Normal tissue homogenate | 1:5, 1:10, 1:20 | Collagen content depends on tissue type (bone > skin > tendon > organ parenchyma) |
| Fibrotic tissue homogenate | 1:20, 1:50, 1:100 or higher | Pathological ECM deposition can elevate Col-I by several-fold to >10-fold over baseline |
| Cell culture supernatant | Neat to 1:10 | Dependent on cell type and culture conditions |
Evaluation Criteria: Choose the dilution that yields an OD value falling within the 5–20 ng/mL mid-range of the standard curve, where precision is highest. Calculate recovery and linearity across dilutions to validate your chosen factor.
Critical Dilution Factor
The assay includes a 5-fold in-well dilution: 10 µL sample + 40 µL Sample Diluent. The concentration interpolated from the standard curve represents this diluted sample. Multiply the interpolated result by 5 and by any additional pre-dilution factor to obtain the concentration in the original specimen.
Example: If a serum sample pre-diluted 1:4 gives an interpolated reading of 8 ng/mL, the actual concentration = 8 × 5 × 4 = 160 ng/mL.
Technical Specifications
| Parameter | Specification |
|---|---|
| Standard Concentrations (S0–S5) | 0, 2.5, 5, 10, 20, 40 ng/mL |
| Quantitative Range | 1.0 – 40 ng/mL |
| Analytical Sensitivity | < 1.0 ng/mL |
| Accuracy (Linearity) | Standard curve R ≥ 0.9900 |
| Precision | Intra-assay CV < 15%; Inter-assay CV < 15% |
| Specificity | Highly specific for native rabbit Type I collagen. No significant cross-reactivity with other collagen types (II, III, IV) or ECM structural analogues. |
| Species Restriction | Validated exclusively for rabbit samples. |
| Sample Dilution Factor | 5× (in-well); additional pre-dilution required |
| Shelf Life | 6 months |
| Storage & Transport | 2–8°C, protected from light and moisture; cold-chain shipping |
Representative Standard Curve
Detailed performance data, including spike-recovery and dilution linearity, are available in the product instruction manual.
Kit Components
96-Well Format
| Component | Quantity | Notes |
|---|---|---|
| Microelisa Stripplate | 12 strips × 8 wells | Pre-coated with anti-rabbit Col-I antibody |
| Standard (0.3 mL/vial) | 6 vials | Ready-to-use liquid, 0–40 ng/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 |
| 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 during cold storage is normal; warm in a water bath until crystals dissolve completely before dilution.
Assay Procedure Summary
- Equilibrate: Warm the sealed foil pouch to room temperature for 20 min. Remove required strips; reseal unused strips at 4°C.
- Add Standards & Samples:
- Standard wells: 50 µL of each standard.
- Sample wells: 10 µL of pre-diluted sample + 40 µL Sample Diluent.
- Blank well(s): Leave empty.
- Add Detection Antibody: Dispense 100 µL HRP-conjugated detection antibody into all wells except blank. Seal and incubate at 37°C for 60 min.
- Wash: Aspirate, blot dry. Fill wells with 1× Wash Buffer, soak 1 min, discard. Repeat 5 times (automated: 350 µL/well, 1-min soak).
- Substrate: Add 50 µL each of Substrate A and Substrate B. Incubate in the dark at 37°C for 15 min.
- Stop & Read: Add 50 µL Stop Solution. Read absorbance at 450 nm within 15 min.
Total hands-on time: ~2 hours.
Materials Required but Not Supplied:
- Microplate reader (450 nm)
- Adjustable pipettes (0.5–1000 µL) and tips
- 37°C incubator or water bath
- Distilled/deionized water
- Absorbent paper, wash bottle or automated washer
- Pepsin and acetic acid (optional, for cross-linked collagen extraction)
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 accordingly.
Related Products
- Rabbit Type III Collagen (Col‑III) ELISA Kit — For fibrosis, vascular and wound healing studies
- Rabbit Type II Collagen (Col‑II) ELISA Kit — For cartilage and osteoarthritis research
- Rabbit Immunoglobulin G (IgG) ELISA Kit – Reliable Detection of Secondary Immune Response
- Rabbit Thyroxine (T4) ELISA Kit – Accurate Thyroid Hormone Quantification
Frequently Asked Questions
Q: Is this a total collagen assay?
A: No. This kit specifically detects Type I collagen, not total collagen. For total collagen, please refer to our human total collagen kit (Yanda-HUMAN-Col).
Q: Does this kit cross-react with rabbit Type II or Type III collagen?
A: No significant cross-reactivity has been observed with other collagen types. The kit is highly specific for rabbit Type I collagen.
Q: Can I use standard PBS homogenization for fibrotic tissues?
A: Yes, PBS homogenization is the standard method. However, highly cross-linked mature collagen in advanced fibrosis may be poorly extracted by PBS alone. In such cases, consider adding a pepsin digestion step. We recommend comparing both methods in a pilot experiment.
Q: What pre-dilution should I start with?
A: For serum/plasma, start with 1:2, 1:4, and 1:8. For tissue homogenates, try 1:5, 1:10, and 1:20. For fibrotic tissues, include 1:50 and 1:100. Choose the dilution that gives an OD in the 5–20 ng/mL mid-range.
Q: How do I calculate the final concentration?
A: Final concentration (ng/mL) = Interpolated value × 5 (in-well dilution) × pre-dilution factor. Example: 1:4 pre-dilution, interpolated reading 8 ng/mL → 8 × 5 × 4 = 160 ng/mL.
Q: Can I use this kit for mouse or human Col-I?
A: No. This kit is validated only for rabbit samples. Species-specific kits for other models may be available upon request.
Q: Does this kit detect recombinant Col-I?
A: The kit is designed to detect native Col-I. Performance with recombinant protein has not been fully characterized.
Q: Are MSDS and composition documents available for export?
A: Yes, these documents can be provided upon request. The kit ships in a neutral white box with cold-chain packaging.
Ordering & Contact Information
| Item | Details |
|---|---|
| Catalog Number | Yanda-Rabbit-Col-I |
| 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 species-specific collagen kits, please reach out directly.
Technical Documents & Support
- MSDS & Certificate of Composition – Available upon request.
- Technical Support – For tissue extraction protocols, dilution optimization, or validation data, contact us via WhatsApp or email.
Disclaimer
This product is for laboratory research use only and is not approved for clinical diagnosis, treatment, or prognosis. The manufacturer assumes no liability for improper use, protocol deviations, or unauthorized applications. Users are responsible for validating assay performance in their own sample matrices and experimental conditions.
