Easy to Use but Inconsistent Replicates

April 15, 2021

Mayo Clinic
Vaccine Research
Lab Technician

Overall

Quality of Results

Ease-of-Optimization

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Company:

R&D Systems

Product Name:

Human IFN-Gamma ESLISpot

Catalog Number:

EL285

We studied IFN-Gamma secretions from T cells using isolated T cells from whole PBMC's in these ELISpot kits.

Experimental Design and Results Summary

Application

Assessing IFN-Gamma Secretions from T cells using whole PBMC's

Starting Material

N/A

Tips

Ensure pipette calibration before plating cells. If more than one operator, eliminate all differences in plating/pipetting technique.

Results Summary

Our optimization was conducted using two operators plating cells from the same subjects. We saw interoperator differences in spot counts as well as interassay differences. Several iterations of optimization was conducted to attempt to correct these differences. Even when one operator assayed two plates, there will still differences between the plates, making it difficult to accurately determine spot counts.

DOI or PMID #

N/A

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Summary

The Good

Packaging, clear protocol,

The Bad

Hard to produce consistent results

The Bottom Line

ELISpots are a variable assay, so it cannot be determined if the kit was at fault for the differences we witnessed. It is best to use whole PBMC's to preserve cell viability without doing any isolations before with this kit. Other kits from R&D have performed well in the past. Optimize with diligence and there should not be an issue.

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