Target Distinction — HMW vs. Total Adiponectin
This kit specifically measures high molecular weight (HMW) adiponectin, the 12–18‑mer multimeric form that is the primary biologically active isoform responsible for insulin sensitization and anti‑inflammatory effects. It does not detect low molecular weight (LMW) or medium molecular weight (MMW) adiponectin.
If you need to measure all adiponectin isoforms simultaneously, please refer to our Human Total Adiponectin ELISA Kit (Yanda-Human-ADPN) . The two kits can be used together to calculate the clinically informative HMW/Total adiponectin ratio.
Key Features
- Direct HMW Detection — No Protease Pretreatment Required: Utilizes monoclonal antibodies that recognize a unique conformational epitope present only on HMW adiponectin. No proteinase K digestion or separation step is needed.
- Clear Differentiation from Total Adiponectin Kit: The HMW kit has a lower detection range (3.9–250 ng/mL vs. 50–1600 ng/mL) and higher sensitivity (<0.989 ng/mL), optimized for the 30–50% fraction of total adiponectin that is HMW.
- Optimized Pre‑Dilution for Serum: Normal serum HMW adiponectin is ~1–10 μg/mL. A pre‑dilution of 1:50 to 1:200 is recommended — less than the 1:100–1:500 typical for total adiponectin, reducing matrix effects.
- Built‑in 5× Sample Dilution: 10 μL sample + 40 μL Sample Diluent in‑well; multiply the interpolated result by 5 to obtain the concentration in the diluted sample.
- Multimer Stability Guidance: HMW adiponectin is sensitive to dissociation by freeze‑thaw, extreme pH, or denaturants. The protocol advises single‑use aliquots, addition of 5–10% glycerol for long‑term storage, and avoidance of repeated freeze‑thaw cycles.
- Ready‑to‑Use Liquid Standards: Six standards (0, 15.625, 31.25, 62.5, 125, 250 ng/mL) supplied as liquids; no reconstitution required.
- 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 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
Adiponectin circulates in three major oligomeric forms: low molecular weight (LMW, trimer), medium molecular weight (MMW, hexamer), and high molecular weight (HMW, 12–18‑mer). Among these, HMW adiponectin is the most metabolically active, exhibiting potent insulin‑sensitizing, anti‑inflammatory, and cardioprotective properties. Clinical and experimental studies consistently show that HMW adiponectin, and particularly the HMW/Total adiponectin ratio, correlates more strongly with insulin sensitivity, type 2 diabetes risk, and cardiovascular outcomes than total adiponectin alone.
The Yanda-Human-HMWADPN ELISA Kit uses a one‑step double‑antibody sandwich ELISA with antibodies specific for the HMW conformational epitope. It provides a direct, digestion‑free method for quantifying HMW adiponectin in serum, plasma, cell culture supernatant, and tissue homogenates.
Sample Collection & Preparation
Sample Types
| Sample Type | Collection & Processing |
|---|---|
| Serum | Collect in pyrogen/endotoxin‑free tubes. 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 debris. |
| Tissue Homogenate | Homogenize tissue in ice‑cold PBS (1:9 w/v) with protease inhibitors. Centrifuge at 3000 rpm for 10 min at 4°C. |
Pre‑Dilution Recommendation
Normal human serum HMW adiponectin is approximately 1–10 μg/mL (1000–10,000 ng/mL). The kit’s quantitative range is 3.9–250 ng/mL. We recommend a pre‑dilution of 1:50 to 1:200 with the provided Sample Diluent (e.g., 2 μL serum + 98 μL diluent for 1:50). Run a pilot gradient (1:50, 1:100, 1:200) to determine the optimal factor for your population.
Multimer Stability & Storage
- Avoid repeated freeze‑thaw cycles. Aliquot serum/plasma immediately after collection and store at -20°C or -80°C.
- For long‑term storage, add 5–10% glycerol to the aliquot to stabilize HMW conformation.
- Do not use reducing agents (e.g., DTT, β‑mercaptoethanol) or denaturing detergents (e.g., SDS >0.1%) in sample preparation, as they can dissociate HMW multimers.
- NaN₃ Prohibition: Sodium azide is an HRP inhibitor and must be absent from samples.
Dilution & Calculation
- In‑well dilution: 10 μL pre‑diluted sample + 40 μL Sample Diluent = 5‑fold dilution.
- Interpolated value: Concentration read from the standard curve (in‑well concentration).
- Final sample concentration (ng/mL) = Interpolated value × 5 (in‑well) × pre‑dilution factor.
Example: Serum pre‑diluted 1:100 gives an interpolated reading of 50 ng/mL. Actual HMW concentration = 50 × 5 × 100 = 25,000 ng/mL = 25 μg/mL.
Technical Specifications
| Parameter | Specification |
|---|---|
| Standard Concentrations (S0–S5) | 0, 15.625, 31.25, 62.5, 125, 250 ng/mL |
| Quantitative Range | 3.9 – 250 ng/mL |
| Analytical Sensitivity | < 0.989 ng/mL |
| Accuracy (Linearity) | Standard curve R ≥0.9900 |
| Precision | Intra‑assay CV <15%; Inter‑assay CV <15% |
| Specificity | Highly specific for human HMW adiponectin via a unique conformational epitope. No cross‑reactivity with LMW or MMW adiponectin, or other soluble structural analogues. |
| Species Restriction | Validated for human samples only. |
| Sample Dilution Factor | 5× (in‑well); additional pre‑dilution of 1:50–1:200 recommended |
| Shelf Life | 6 months |
| Storage & Transport | 2–8°C, protected from light; cold‑chain shipping |
Representative Standard Curve
Detailed performance 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‑HMW adiponectin antibody |
| Standard (0.3 mL/vial) | 6 vials | Ready‑to‑use liquid, 0–250 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 storing unused strips |
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 (Recommended for Cross‑Species Studies)
Expand your metabolic research with these complementary kits (available from Yanda Bio):
- Rat Adiponectin ELISA Kit – for rodent models of obesity, diabetes, and metabolic syndrome
- Human Adiponectin (ADPN) ELISA Kit—High‑Sensitivity Sandwich Assay
- Quantify Human 2,3-DPG ELISA Kit with a Superior Signal-to-Noise Ratio.
- Human Histidine (HIS) ELISA Detection Kit – Competitive Assay for Amino Acid Quantification
Using human, rat and mouse adiponectin kits together allows cross‑species comparisons and validates translational findings.
Frequently Asked Questions
Q: Does this kit detect total adiponectin or only HMW?
A: Only HMW adiponectin. The antibodies recognize a unique conformational epitope present on the 12–18‑mer complex, with no cross‑reactivity to LMW or MMW forms.
Q: Do I need to pretreat samples with proteinase K?
A: No. The kit directly detects HMW adiponectin without any digestion or separation step.
Q: How is this different from the Human Total Adiponectin Kit (Yanda-Human-ADPN)?
A: The Total kit measures all isoforms (LMW+MMW+HMW) with a range of 50–1600 ng/mL. This HMW kit specifically measures the HMW fraction with a range of 3.9–250 ng/mL. Use both together to calculate the HMW/Total ratio.
Q: What pre‑dilution should I use for serum?
A: Normal HMW adiponectin is ~1–10 μg/mL. A pre‑dilution of 1:50 to 1:200 is recommended. Perform a pilot gradient to find the optimal factor.
Q: How can I calculate the HMW/Total ratio?
A: Measure the same sample with both the HMW kit (this product) and the Total Adiponectin kit (Yanda-Human-ADPN). Divide the HMW concentration by the Total concentration. The ratio is a superior marker of metabolic health than either parameter alone.
Q: How do I prevent HMW from dissociating during storage?
A: Avoid repeated freeze‑thaw cycles. Aliquot samples and add 5–10% glycerol as a stabilizer. Do not use reducing agents or high SDS concentrations.
Q: Can I use this kit for mouse or rat samples?
A: No. This kit is validated for human samples only. Use species‑specific kits for rodent HMW adiponectin.
Q: Are MSDS and export documents available?
A: Yes. MSDS and Certificate of Composition are provided upon request.
Ordering & Contact
| Item | Details |
|---|---|
| Catalog Number | Yanda‑Human‑HMWADPN |
| 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 HMW/Total ratio calculation, multimer stability, or validation data, 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.
