EnzyChrom™ Gliadin ELISA Kit

The background is very high?

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.

Can I store the diluted standards (0ppb, 1.2ppb, 0.6ppb, 0ppb) to use for later?

We recommend preparing the gliadin standard fresh from the 2000ppb stock before each assay.

The TMB Substrate is frozen after being stored at 4°C and taking very long to thaw?

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.

I don't know how much gluten/gliadin is in my sample. How much should I dilute it?

If the gluten/gliadin level is unkown, we recommend running a serial dilution on the Sample Extract to determine the necessary dilution. If you have an estimate of the gluten level (e.g. very high, low, or very low), the dilution guidelines table in the protocol can serve as a decent starting point.

There is a fair amount of variation in my assay?

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.

The calculated gliadin for my samples seems to be much too high/low?

Because the assay is very sensitive, it typically requires very large dilution factors. The samples may need to be greater diluted before assay or may be diluted too much. For unknown samples, we recommend running a titration of dilutions. Your ideal dilution should produce an OD in the middle of the standard curve.

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

Yes.

My sample has barley and/or rye in it, but is showing up as not having any gliadin?

This assay only quantifies gliadin, the predominant prolamin in wheat. Barley and rye contain different prolamins than gliadin.

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.

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 5× 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.