EnzyChrom™ Peanut ELISA Kit
Protocol MSDS


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Application
•   Determination of peanut in food and beverage samples.

Key Features
•   Fast and Simple. Entire assay procedure takes less than three hours.

•   Sensitive and Accurate. Quantifies peanut in the parts per billion range.

•   Simple Calculation. Simple calculation with linear standard curve.

Method
•  Sandwich ELISA (OD450nm)

Samples
•   Food and beverage samples

Species
•   All

Procedure
•   3 hours

Size
•   96 tests

Detection Limit
•   5 ppb

Shelf Life
•   12 months

More Details
•   Peanut allergy is one of the most common and most dangerous food allergies. Trace amounts of peanut can be enough to trigger life-threatening anaphylaxis, which if left untreated, can result in death. Due to the severity and prevalence of peanut allergies, regulation of peanut containing foods is strictly enforced. The allergens Ara h1, Ara h2, Ara h3, and Ara h6 are the most prominent peanut allergens that cause allergic reactions. The protein Ara h3 is heat stable and comprises a significant portion of the protein in peanuts. These traits make Ara h3 an ideal candidate for quantifying peanut content in samples, even those that have been cooked or baked.
BioAssay Systems' Peanut ELISA uses a colorimetric sandwich ELISA method specific for the protein Ara h3 to quantify the peanut content of samples.

1. What may be the issue when I have a high background?



This can be caused by residual detection antibody being left in the well. Please make sure to properly and thoroughly wash the wells for the final wash step. If there is residual wash buffer in the wells, dump it out.

2. Can I store the diluted standards (50, 30, 15, 0 ppb) to use for later?



We recommend preparing the peanut standards fresh from the 10 ppm stock before each assay.

3. The TMB Substrate is frozen after being stored at 4°C and taking very long to thaw. Is this an issue?



The TMB Substrate is in the solvent DMSO, which has a freezing point of 19°C. Please be sure to allow adequate time to thaw before using in the assay.

4. I don't know how much peanut is in my sample. How much should I dilute it?



If the peanut level is unknown, we recommend running a serial dilution on the Sample Extract to determine the necessary dilution. If you have an estimate of the peanut content, the dilution guidelines table in the protocol can serve as a starting point.

5. There is a fair amount of variation in my assay, how can I fix this?



Ensure that the washing technique being used is efficient. If an automated plate washer is available, consider using that. ELISAs have many steps and thus many points for errors to be introduced. Be sure all pipetting is precise and accurate. Use of a multichannel pipettor is recommended if available.

6. The calculated peanut for my samples seems to be much too high/low. Why may this be happening?



Because the assay is very sensitive, it typically requires very large dilution factors. The samples may have been over diluted or not diluted enough. For unknown samples, we recommend running a titration of dilutions. Your ideal dilution should produce an OD in the middle of the standard curve.

7. Do I need to run a standard curve with every assay?



Yes, it is prudent to run a standard curve with each assay

8. Is this kit approved by the FDA?



The kit is intended for research purposes only. It is not approved by the FDA for food analysis or quality control.

9. I don't have a 60 °C waterbath, can I heat the samples at 100°C?



No, heating the samples at 100°C will destroy the peanut protein rendering it incompatible with the ELISA.

10. My sample is limited. I don't have 1 g or 1 mL to use for sample extraction. What can I do?



If there is not enough sample to use 1 g or 1 mL in the extraction, you may use a smaller weight or volume. Please be sure to note the different w/v or v/v concentration when correcting for dilutions in the calculation. For instance, 0.5 g or 0.5 mL diluted to 10 mL would be a 5% w/v or v/v solution and 0.1 g or 0.1 mL diluted to 10 mL would be a 1% w/v or v/v solution.

11. How should I store the kit?



The kit is shipped on ice. Upon delivery, store Detection Ab and Standard at -20°C. Store all other reagents at 4°C. Note: all reagents may be stored at -20°C; however, precipitates may form in 10× Wash Buffer and 10× Sample Buffer. If precipitates form, heat in a hot water bath until precipitates are solubilized before using.

For more detailed product information and questions, please feel free to Contact Us. Or for more general information regarding our assays, please refer to our General Questions
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Please inquire or request assay service or call 1-510-782-9988 x 2.
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EnzyChrom™ Peanut ELISA Kit
Catalog No: EPNT-100
Price: $509    Qty:
For orders of 10 or more kits, please call 1-510-7829988x1 or email us for best pricing and/or bulk order.

Shipping: On Ice
Shipment: Fedex Service
Delivery: 1-2 days (US), 3-6 days (Intl) Storage: -20°C

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