Human ELISA kit
Human 1,3-Beta-D-glucan (BDG) ELISA Kit
Frequently Asked Questions
How to ensure that the ELISA kit will not be damaged during transportation.
We will use air transportation to ensure that the test kits are delivered to you within one week. During this period, we used EPS foam boxes to store the ELISA kits and used dry ice to ensure the temperature was below 20°C.
Could you possibly offer a trial pack?
If you need antibodies, we can offer a 10-microliter trial pack for you, but you will be responsible for the shipping cost. We do not provide the ELISA kit packaging, but we can offer you an ELISA kit at a price much lower than the market rate for your use.
What Payment Methods Do You Support?
Currently, we support credit card payment, bank transfer, and third-party payment platforms (such as Alipay,Paypal.WeChat Pay, etc. International business will be adjusted according to relevant regulations).
Could you offer ELISA testing service for me?
Yes.If you purchase our test kits, we will offer a free ELISA testing service. If you provide the test kits yourself, we will charge a testing fee of $50.
Could you offer the service of customizing ELISA kits?
Yes, we offer custom ELISA kit services. Both the test factors and the detection sensitivity of the ELISA kits can be customized.
Why is your product so cheap?
We are the original manufacturer and this is our first attempt to enter the international market. We hope to establish trust with you as quickly as possible. Believe me, choosing to trust us will be the best decision for you.
Operational Skills in ELISA Experiments
In China, we provide thousands of ELISA test kits to researchers every year. Occasionally, there are cases where customers are unable to detect the test factors. When such situations occur, we offer a testing service for the researchers. After testing with the same ELISA kits using the customers’ samples, we found that all the test factors could be detected. Therefore, we suspect that this situation may be related to the customers’ operations.Carry out the experiment strictly in accordance with the operation procedures specified in the ELISA kit manual, which will ensure the most accurate test results.
Tip 1: Wipe the Plate Bottom After Enzyme Addition
After adding enzyme conjugate, gently wipe the bottom of the microplate with absorbent paper to remove dust, fingerprints, or liquid droplets.
Why it matters: Even minor residues on the plate bottom can scatter or absorb light, affecting optical density (OD) readings. A clean plate bottom reduces well‑to‑well variability by 5–10% and ensures that measured OD reflects only the colorimetric reaction .
Tip 2: Plan Your Plate Layout to Minimize Delays
Enzyme-linked immunosorbent assay (ELISA) remains the gold standard for protein quantification in life science research. Its popularity stems from high sensitivity, strong specificity, excellent reproducibility, and relatively simple operation. Reagents are stable, easy to store, and results can be interpreted objectively using a standard microplate reader.
However, even experienced researchers encounter variability that compromises data quality. The difference between publication-ready results and frustrating inconsistencies often lies in the subtle techniques applied during execution.
This guide presents 15 evidence-based tips to optimize your ELISA workflow—from sample handling to data interpretation—with supporting data to help you achieve maximum accuracy and reproducibility.
When working with multiple plates, calculate your sample count in advance and arrange wells to avoid prolonged waiting times between steps.
The problem: If you have 5 plates but only one incubator, the last plate may sit at room temperature 10–15 minutes longer than the first—enough time for nonspecific binding to increase background by 0.05–0.10 OD units .
Solution: Process plates in batches that match your incubator capacity and pipetting speed. For 96‑well plates, limit batches to 2–3 plates at a time.
Tip 3: Use Volume-Matched Pipettes
Always select a pipette with a volume range closest to the volume you’re dispensing.
For 10 µL, use a 2–20 µL pipette, not a 100–1000 µL pipette.
For 100 µL, use a 20–200 µL pipette.
Data point: A 1000 µL pipette set to 100 µL has an accuracy of ±3.0%, while a 200 µL pipette at the same volume achieves ±0.8% accuracy . This 2.2% difference translates to significant well‑to‑well variation across a 96‑well plate.
Tip 4: Always Run Duplicate Wells
Running samples in duplicate (or triplicate) is not optional—it’s essential for data integrity.
Duplicate wells allow calculation of the coefficient of variation (CV), which should be <10% for reliable results.
Single wells cannot distinguish between experimental variation and pipetting errors.
Real-world impact: In a study comparing single vs. duplicate wells, single-well measurements deviated from true values by an average of 12.3% , while duplicates reduced this error to 4.1% .
Tip 5: Calibrate Your Sample Diluter Regularly
If using an automated diluter for sample preparation, verify its accuracy monthly.
The risk: A mis-calibrated diluter dispensing 90 µL instead of 100 µL introduces a 10% systematic error across all samples—enough to shift results outside the linear range of your standard curve.
Quick check: Weigh dispensed water (1 g = 1 mL) to confirm volume accuracy.
Tip 6: Prevent Evaporation During Incubation
Evaporation is a silent killer of ELISA accuracy. To minimize it:
Place plates in a humidified box lined with wet paper towels
Always seal plates with adhesive film or cover lids
Data: Unsealed plates lose 5–10 µL/well per hour at 37°C . Over a 2‑hour incubation, this concentration effect can artificially increase OD by 15–20% , leading to false positives or overestimation.
Tip 7: Store Unused Strips and Reagents Properly
Return unused strips to the resealable bag with desiccant and store at 2–8°C
Prepare only the needed amount of HRP-conjugated working solution—do not reuse diluted conjugate
Why: Diluted HRP conjugate loses 30–50% of its activity within 24 hours at 4°C . Reusing it leads to signal decay and inconsistent results.
Tip 8: Batch Samples to Control Incubation Timing
When processing many samples, add reagents in the same order and timing for each batch.
The risk: If you take 20 minutes to add samples to 96 wells, the first well incubates 20 minutes longer than the last. This can increase background by 0.08–0.12 OD in the first wells .
Solution: Use a multichannel pipette and limit batch size to 32 wells (one strip) at a time.
Tip 9: Dispose of Biohazardous Waste Properly
All samples and contaminated materials must be inactivated before disposal:
Autoclave at 121°C for 30 minutes
Or treat with 5.0 g/L sodium hypochlorite for 30 minutes
Compliance note: Proper disposal is not just safety—it’s often required for institutional animal care and use protocols.
Tip 10: Fill Wells Completely During Washing
When washing manually or with an automated washer, ensure each well is filled to the brim (300–350 µL for 96‑well plates).
Why it matters: Incomplete filling leaves unbound enzyme at the well top, which can trickle down during subsequent steps and increase background by 0.10–0.15 OD .
Tip 11: Reserve One Well for Blank Subtraction
Always include a blank well that receives:
No sample
No detection antibody
Only substrate and stop solution
Procedure: After adding stop solution, use this well to zero your plate reader. This corrects for plate imperfections and reagent background.
Impact: Blank subtraction typically reduces background by 0.03–0.07 OD , improving signal‑to‑noise ratio by 10–15% .
Tip 12: Master Manual Washing Technique
If washing by hand:
Add wash buffer and let sit 15–30 seconds per cycle
Avoid splashing between wells (causes cross‑contamination)
After aspiration, firmly tap plate on absorbent paper 3–5 times
Data: Proper manual washing achieves residual volumes of 2–5 µL/well , comparable to automated washers. Poor technique leaves 15–20 µL , increasing background by 0.12–0.18 OD .
Tip 13: Validate Questionable Results
If a sample yields unexpected values (too high, too low, or inconsistent), verify using an orthogonal method such as Western blot or LC‑MS/MS.
Why: No single assay is perfect. Cross‑validation catches false positives/negatives caused by sample matrix effects or antibody cross‑reactivity.
Tip 14: Use High-Quality Water
When deionized or distilled water is unavailable, Wahaha purified water (or equivalent brand) is acceptable. Never use tap water.
The risk: Tap water contains ions and organic compounds that can:
Alter pH of buffers
Introduce enzyme inhibitors
Increase background by 0.15–0.25 OD
Tip 15: Change Tips Between Reagents
This seems obvious, but it’s a common source of contamination. Always use a fresh tip when moving between different reagent bottles.
Critical point: When preparing standard dilutions, change tips between each dilution step to avoid carryover that can skew your curve by 10–20% .
Bonus Tip: Avoid Air Bubbles
When pipetting, dispense slowly to prevent bubble formation. Bubbles in wells:
Interfere with antibody binding
Cause variable light path length during reading
Increase well‑to‑well CV by 5–8%
If bubbles appear, gently tap the plate or burst them with a clean needle before reading.
Summary: Small Changes, Big Impact
| Technique | Potential Improvement |
| Wipe plate bottom | 5–10% lower variability |
| Use volume‑matched pipettes | 2.2% better accuracy |
| Run duplicates | 8% reduction in error |
| Prevent evaporation | 15–20% less signal drift |
| Proper washing | 0.12–0.18 OD lower background |
Why Choose Yanda Bio for Your ELISA Needs?
At Yanda Bio, we don’t just sell kits—we partner with researchers to ensure experimental success. Our 6,000+ ELISA kits cover immunology, oncology, neuroscience, metabolism, and infectious diseases, with sensitivities customizable down to 60 pg/mL.
What sets us apart:
Rigorous validation: Every lot tested for precision (CV <10%) and accuracy
Affordable pricing: Standard kits from just $120
Free ELISA testing service: Buy kits, send samples—we test for you
Rapid delivery: Same‑day dispatch for orders before 3:30 PM
Technical support: Expert troubleshooting when you need it