Biomarker ELISA Kits

ACRO kits are validated based on ICH M10 guidelines, with performance data (inter-assay/intra-assay precision, dilution linearity, recovery) included in the method validation report. To obtain the complete report, contact ACRO Technical Support.

When available, our standards are anchored to NIBSC/WHO international reference materials. Correction factors are provided in the product webpage. If you have any questions, refer to the product webpage or contact technical support.

Our kits are validated to tolerate interference from:
• Cell culture media（X-VIVO, DMEM etc.）
• Hemoglobin (2% v/v)
• Triglycerides (≤3 mg/mL)
• Bilirubin (≤20 mg/dL)
Full validation data is available in the method validation reports, please contact ACRO sales and technical support.

For information of kit components, please contact ACRO sales and technical support.

Yes, our kits are validated using healthy human serum/cell supernatant specimens. Validation includes:
• Healthy donor serum/cell supernatant (n=30 minimum)
• Spike-and-recovery testing in clinical matrices (≥80% recovery required)

The four-parameter logistic (4PL) equation is recommended, as it is the most widely used fitting method for immunoassays internationally. The 4PL equation is suitable for various curve shapes, including monotonic curves (exponential, logarithmic, hyperbolic) and sigmoidal curves. Please notice that the X value must not be less than 0 (since exponents are real numbers).

The ULOQ OD value for ELISA kit standard curve is typically between 2.0 and 2.5. Slight variations may occur due to differences in the real laboratory conditions.

Configure the instrument as follows:
• Measurement Mode: Absorbance (Optical Density, OD)
• Primary Wavelength:450 nm (optimized for the kit’s substrate solution)
• Reference Wavelength: 630 nm

Included Components:

• Pre-coated microplate
• Target-specific antibodies
• Calibrated protein standards
• Streptavidin-HRP conjugate
• TMB substrate solution
• Wash buffer (10× concentrate)
• Stop solution

User-Supplied Materials:

• Pipettes & tips (multi-channel recommended)
• Centrifuge tubes
• Ultrapure water

For full component specifications, see the Components List in the manual.

Critical Steps:

1. Reagent Handling:

o Thaw all components at 20–25°C for 30 mins
o If crystals form in concentrated wash buffer, warm at 37°C for 5 mins then mix gently

2. Lyophilized Standard Reconstitution:

o Reconstitute lyophilized components with ultrapure water
o Mix components by pipetting 5-10 times, rest for at least 15 mins (normally 15 – 30 mins) before use.

3. Equipment Check:

o Validate pipette calibration
o Confirm microplate reader warm-up (≥15 mins)

• Optimal Condition: 22–25°C (ambient lab environment)
• Risk of 37°C:

* Accelerated HRP activity → Premature substrate exhaustion

Three critical timing controls:

1. Follow incubation times strictly.
2. Terminate substrate reaction immediately when all wells turn uniform yellow (10–15 minutes).
3. Read results within 10 minutes after adding stop solution.

For detailed procedures and precautions, refer to the product manual or consult ACRO technical support.

Not recommended: Substitutions may compromise results.

Exceptions: Contact ACRO technical support before use for approval.

1. Unopened: Store according to the expiration date on the label (e.g., 2–8°C).

2. Opened:

• Reconstituted standards: -80°C, use within 1 month.
• Substrate solution: 4°C (dark), use within 1-2 days.