Aß Peptide Measurements In Human Biofluids

November 01, 2016

University of Southern California
Physiology and Biophysics
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

What do these ratings mean?
Write a Review

Company:

Meso Scale Discovery (MSD)

Product Name:

V-PLEX Aβ Peptide Panel 1 (6E10) Kit

Catalog Number:

K15200E-1

I used this kit to detect Aβ peptide (38, 40, and 42) 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: cerebrospinal fluid (CSF) and plasma

Protocol Overview

The 96-well plate is blocked for 1 hr at room temperature with blocking solution. Meanwhile the diluted samples and standard are prepared. Then, the plate is washed with wash buffer. The prepared antibody is added to each well, and 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. Then 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 an orbital 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.

Image Gallery

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.

Join the discussion