Introduction
Alterations in the levels of vascular inflammation proteins have been associated with cardiovascular disease, cancer, rheumatoid arthritis, kidney function,
bacterial or viral infection, and liver failure. Accurate quantification of proteinaceous biomarkers involved in these inflammatory and immune disorders is an
area of ongoing biomedical research, and important to further understanding
these disease states.
The Human Vascular Inflammation Panel 1 is a multiplex bead-based assay
panel, using fluorescence–encoded beads suitable for use on various flow cytometers. This panel allows simultaneous quantification of 13 human proteins,
including Myoglobin, Calprotectin (MRP8/14), Lipocalin A (NGAL), C-Reactive
Protein (CRP), MMP-2, Osteopontin (OPN), Myeloperoxidase (MPO), Serum Amyloid A (SAA), IGFBP-4, ICAM-1 (CD54), VCAM-1 (CD106), MMP-9, and Cystatin
C. This assay panel provides higher detection sensitivities and broader dynamic
ranges than traditional ELISA methods. The panel has been validated for use
with serum (S), plasma (P), urine (U) and tissue culture supernatants (TC).
The LEGENDplexTM Human Vascular Inflammation Panel 1 is designed to allow
flexible customization within the panel. It can also be divided into sub-panels
based on biological sample type and suggested dilution factor, listed below:
LEGENDplexTM Human Vascular Inflammation Panel 1-TC (13-plex)
LEGENDplexTM Human Vascular Inflammation Panel 1-S/P (12-plex)
LEGENDplexTM Human Vascular Inflammation Panel 1-CRP (1-plex)
LEGENDplexTM Human Vascular Inflammation Panel 1-U (10-plex)
LEGENDplexTM Human Vascular Inflammation Panel 1-U (3-plex)
Please visit www.biolegend.com/legendplex for more information on panel
design and how to mix and match within the panel.
Principle of the Assay
BioLegend’s LEGENDplexTM assays are bead-based immunoassays using the
same basic principle as sandwich immunoassays.
Beads are differentiated by size and internal fluorescence intensities. Each bead
set is conjugated with a specific antibody on its surface and serves as the capture beads for that particular analyte. When a selected panel of capture beads
is mixed and incubated with a sample containing target analytes specific to the
capture antibodies, each analyte will bind to its specific capture beads. After
washing, a biotinylated detection antibody cocktail is added, and each detection antibody in the cocktail will bind to its specific analyte bound on the capture beads, thus forming capture bead-analyte-detection antibody sandwiches.
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LEGENDplex™ Human Vascular Inflammation Panel 1
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Streptavidin-phycoerythrin (SA-PE) is subsequently added, which will bind to
the biotinylated detection antibodies, providing fluorescent signal intensities in
proportion to the amount of bound analytes.
Since the beads are differentiated by size and internal fluorescence intensity
on a flow cytometer, analyte-specific populations can be segregated and PE
fluorescent signal quantified. The concentration of a particular analyte is determined using a standard curve generated in the same assay