Key Features
- Detects Rat Total Adiponectin – Measures all multimeric forms (LMW, MMW, HMW) in rat serum, plasma, cell culture supernatant, and tissue homogenates.
- Strict Species Specificity – Validated exclusively for rat samples. No cross‑reactivity with mouse adiponectin. For murine models, a dedicated mouse adiponectin kit is required.
- No Interference from Other Adipokines – No significant cross‑reactivity with leptin, resistin, TNF‑α, or other soluble structural analogues.
- Pre‑Dilution Protocol for High‑Abundance Serum – Normal rat adiponectin levels range from 5–20 μg/mL, far exceeding the assay range. A pre‑dilution of 1:100 to 1:500 is required. A pilot dilution guide is provided.
- Dedicated Adipose Tissue Protocol – Includes mandatory delipidation steps (high‑speed centrifugation or lipid‑clearing agents) and RIPA lysis with protease inhibitors to ensure accurate quantitation from fat‑rich samples.
- Ready‑to‑Use Liquid Standards – Six standards (0, 100, 200, 400, 800, 1600 ng/mL) supplied as liquids; no reconstitution needed.
- Built‑in 5× Sample Dilution – 10 μL sample + 40 μL Sample Diluent in‑well; multiply the interpolated result by 5 and the pre‑dilution factor to obtain the actual concentration.
- One‑Step Sandwich ELISA – Co‑incubation of sample and HRP‑detection antibody; total assay time ~2 hours.
- Excellent Precision – Intra‑ and inter‑assay CV <15%; standard curve 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 and Certificate of Composition available upon request.
Product Description
Adiponectin is a major insulin‑sensitizing adipokine secreted primarily by white adipose tissue. In rats, it plays a critical role in glucose regulation, fatty acid oxidation, and vascular protection. Low circulating adiponectin levels are strongly associated with obesity, insulin resistance, and type‑2 diabetes, making rat models essential for metabolic syndrome research.
The Yanda-Rat-ADPN ELISA Kit uses a one‑step double‑antibody sandwich format. A pre‑coated capture antibody specific for rat adiponectin binds the analyte in standards and samples. An HRP‑conjugated detection antibody is added simultaneously, and after washing, TMB substrate produces a signal directly proportional to adiponectin concentration, measured at 450 nm.
Note: This kit measures total adiponectin. It is validated exclusively for rat samples and does not cross‑react with mouse adiponectin.
Sample Collection & Preparation
Sample Types
| Sample Type | Preparation |
|---|---|
| Serum | Collect without anticoagulant. Allow to clot, centrifuge at 3000 rpm for 10 min. |
| Plasma | Use EDTA, citrate, or heparin. Centrifuge at 3000 rpm for 30 min. |
| Cell Culture Supernatant | Centrifuge at 3000 rpm for 10 min to remove cells and debris. |
| Adipose Tissue | See detailed protocol below. |
Critical Pre‑Dilution for Serum/Plasma
Normal rat adiponectin levels are 5–20 μg/mL (5000–20,000 ng/mL), which is 3–12 times above the kit’s 1600 ng/mL upper limit. Direct measurement without pre‑dilution will saturate the assay.
Recommended pre‑dilution: 1:100 to 1:500. For most samples, a 1:200 dilution (e.g., 2 μL serum + 398 μL Sample Diluent) works well. A pilot gradient (1:100, 1:200, 1:500) is strongly recommended to identify the factor that places OD values in the mid‑range of the standard curve.
Adipose Tissue Preparation
Adipose tissue is rich in lipids that interfere with ELISA.
- Lysis: Homogenize tissue in ice‑cold RIPA buffer (50 mM Tris‑HCl, pH 7.4, 150 mM NaCl, 1% Triton X‑100, 0.5% sodium deoxycholate, 0.1% SDS) supplemented with fresh protease inhibitor cocktail.
- Delipidation: Centrifuge homogenate at 10,000 × g for 10–15 min at 4°C. Carefully aspirate the middle aqueous layer, avoiding the top lipid layer and the pellet. Alternatively, use a commercial lipid‑clearing agent or TCA precipitation (RELi protocol).
- Dilution: The resulting supernatant may still require further pre‑dilution; pilot testing is recommended.
Storage & Stability
- Aliquot and freeze at -20°C or -80°C immediately after collection. Avoid repeated freeze‑thaw cycles.
- NaN₃ Prohibition: Sodium azide inhibits HRP. Do not use as a preservative.
Dilution & Calculation
- In‑well dilution: 10 μL pre‑diluted sample + 40 μL Sample Diluent = 5‑fold dilution.
- Final concentration (ng/mL) = Interpolated value from standard curve (ng/mL) × 5 (in‑well) × pre‑dilution factor.
Example: A serum sample pre‑diluted 1:200 gives an interpolated reading of 80 ng/mL.
Actual adiponectin concentration = 80 × 5 × 200 = 80,000 ng/mL = 80 μg/mL.
Technical Specifications
| Parameter | Specification |
|---|---|
| Standard Concentrations (S0–S5) | 0, 100, 200, 400, 800, 1600 ng/mL |
| Quantitative Range | 50 – 1600 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 rat adiponectin. No cross‑reactivity with mouse adiponectin, leptin, resistin, TNF‑α, or other soluble structural analogues. |
| Species Restriction | Validated exclusively for rat samples. |
| Sample Dilution Factor | 5× (in‑well); additional pre‑dilution of 1:100–1:500 required |
| Shelf Life | 6 months |
| Storage & Transport | 2–8°C, protected from light and moisture; cold‑chain shipping |
Representative Standard Curve
Detailed spike‑recovery and dilution linearity data 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‑rat adiponectin antibody |
| Standard (0.3 mL/vial) | 6 vials | Ready‑to‑use liquid, 0–1600 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 |
| 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.
Assay Procedure Summary
- Equilibrate: Warm the foil pouch to room temperature for 20 min. Remove required strips.
- Add Standards & Samples: 50 μL standards; 10 μL pre‑diluted sample + 40 μL Sample Diluent. Blank: leave empty.
- Add HRP‑Antibody: 100 μL HRP‑detection antibody to all wells except blank. Incubate at 37°C for 60 min.
- Wash: Aspirate, wash with 1× Wash Buffer (soak 1 min), repeat 5 times.
- Substrate: 50 μL each of Substrate A and B, incubate dark at 37°C for 15 min.
- Stop & Read: 50 μL Stop Solution, read at 450 nm within 15 min.
Total hands‑on time: ~2 hours.
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 sample count accordingly.
Related Products
- Rat 3-Indolepropionic Acid (3-IPA) ELISA Kit – Precise Gut Metabolite Quantification
- Rat 5-Alpha Reductase (5AR) ELISA Kit – Precise Enzyme Activity Quantification
- Human Adiponectin (ADPN) ELISA Kit—High‑Sensitivity Sandwich Assay
- Rat 2,3-Disphosphoglycerate (2,3-DPG) ELISA Kit (Competitive ELISA) – For the specific quantitative determination of Rat 2,3-Diphosphoglycerate (2,3-DPG) concentration in hemolyzed erythrocyte samples, serum, and plasma.
Frequently Asked Questions
Q: Does this kit measure total adiponectin or only HMW?
A: Total adiponectin. It detects all multimeric forms (LMW, MMW, HMW) without protease pretreatment.
Q: Can I use this kit for mouse samples?
A: No. This kit is strictly validated for rat adiponectin and does not cross‑react with mouse adiponectin. A dedicated mouse adiponectin kit should be used.
Q: What pre‑dilution should I use for normal rat serum?
A: Normal rat adiponectin is 5–20 μg/mL. We recommend a pre‑dilution of 1:100 to 1:500. Run a pilot gradient to determine the optimal factor.
Q: How do I prepare adipose tissue for adiponectin measurement?
A: Homogenize in RIPA buffer with protease inhibitors, then delipidate by centrifugation (10,000 × g, 15 min) or use a lipid‑clearing agent. Test the aqueous supernatant at several dilutions.
Q: Does this kit cross‑react with leptin or resistin?
A: No. No significant cross‑reactivity with leptin, resistin, TNF‑α, or other adipokines has been observed.
Q: How should I store my samples?
A: Aliquot and freeze at -20°C or -80°C. Avoid repeated freeze‑thaw. Do not use NaN₃.
Ordering & Contact
| Item | Details |
|---|---|
| Catalog Number | Yanda-Rat-ADPN |
| 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 |
WhatsApp: +86 198 3186 0669 | Email: songhawyble@gmail.com
Technical Documents & Support
- Instruction Manual – Download or request via email.
- MSDS & Certificate of Composition – Available upon request.
- Technical Support – For dilution optimization, adipose tissue protocols, or species validation, contact us via WhatsApp or email.
Disclaimer
For research use only. Not for clinical diagnosis or treatment. Users are responsible for validating assay performance in their own experimental conditions. The manufacturer assumes no liability for improper use or protocol deviations.ce to the protocol is required; Yanda Bio is not responsible for results obtained from deviations.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
