Vascular injury markers

February 27, 2015

University of Southern California
Physiology and Biophysics
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Meso Scale Discovery (MSD)

Product Name:

Vascular Injury Panel 2 (human) Kit V-PLEX

Catalog Number:

K15198D

I am interested in detection of molecular biomarkers. I used this kit to detect soluble VCAM-1 and ICAM-1 concentrations in human biofluids (plasma and cerebrospinal fluid). This company, Meso Scale Discovery, specializes in offering multiplex assays with large detection ranges. I am happy with the quality of results obtained from this assay and will continue to purchase from this company. Both the intra- and inter-assay reliability and reproducibility is excellent for this product.

Experimental Design and Results Summary

Application

Quantifying molecular markers

Starting Material

Human biofluids

Protocol Overview

Following desired sample dilutions and standard preparation, the 96-well plate is washed with wash buffer and then the samples/standards are added to wells in duplicate. The plate incubates at room temperature for 2 hours and then is washed a second time. Detection antibody is added to each well and the plate incubates at room temperature for 1 hour. Following a third wash step, read buffer is added to each well and the plate is read on the Meso Scale Discovery multiplex platform via electrochemiluminescence. Concentrations are calculated for each sample from the electrochemiluminescent reading relative to the known standard concentrations.

Tips

I recommend shaking the plate on a microtiter plate shaker during the incubation steps.

Results Summary

The Meso Scale Discovery software "Discovery Workbench" is very user friendly, and the program quickly calculates sample concentrations based on the known standard curve concentrations and dilution of unknown samples.

Additional Notes

Image published in: Montagne A, Barnes SR, Sweeney MD, Halliday MR, Sagare AP, Zhao Z et al. Blood-brain barrier breakdown in the aging human hippocampus. Neuron 2015; 85: 296–302.

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Summary

The Good

The protocol is straight forward and easy to follow.

The Bad

The kits are more expensive than traditional ELISAs

The Bottom Line

Multiplex platforms are advantageous because a single assay can detect 4-10 analytes. This method is ideal over traditional ELISAs if one's model has limited availability of sample volume.

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