FirePlex -HT: A new high- throughput, multiplex …...FirePlex®-HT: A new high- throughput,...
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FirePlex ® -HT: A new high- throughput, multiplex immunoassay ● Performance: ◦ 3-4 logs dynamic range ◦ 1-100 pg/ml sensitivity (*analyte dependent) ● Fast and Automation Friendly: ◦ No-wash assay, Two-step workflow ◦ 6.25 μl of plasma, serum, or cell culture supernatant input ◦ Up to 10 unique analytes per well, and 175 samples in duplicate per 384-well plate ◦ Read out on high-content imagers, <20min scan/plate ● Consistent: ◦ <15% intra-plate CVs ◦ 70-130% sample recovery ◦ Choose from over 800 available antibody pairs ◦ Recombinant pairs, therefore no lot-to-lot variability Assay Workflow: FirePlex ® -HT quantifies up to 10 protein analytes per well from low sample inputs. The two-step workflow and no-wash, 384-well plate assay format limit hands-on time and are amenable to automation, thus making FirePlex ® -HT ideally suited for high-throughput screening studies. Assay readout is conducted on high-content imagers and data analysis is performed with the integrated, free-of-charge FirePlex ® Analysis Workbench software, thus bypassing the need for dedicated instrumentation or expensive software licences. Figure 1. To capture analytes onto FirePlex particles, biological samples are added to a 384-well imaging plate and incubated with the FirePlex-HT Reagent Mix overnight. Subsequently, an Imaging Dye is added and plates are scanned on high-content imagers (refer to Table 1 for a list of currently validated high-content imagers). The FirePlex Analysis Workbench software generates standard curves and quantifies analytes of interest directly from image files. Image Analyze Incubate Samples + Particles + FirePlex ® Reagent Mix Biological samples 1. Shake overnight High-content imager Data analysis with FirePlex ® Analysis Workbench Software Analytes captured onto fluorescently labelled particles 2. Add Imaging Dye 3. Collect Data Key Assay Features:
Transcript of FirePlex -HT: A new high- throughput, multiplex …...FirePlex®-HT: A new high- throughput,...
FirePlex®-HT: A new high-throughput, multiplex immunoassay
● Performance: ◦ 3-4 logs dynamic range
◦ 1-100 pg/ml sensitivity (*analyte dependent)
● Fast and Automation Friendly: ◦ No-wash assay, Two-step workflow
◦ 6.25 μl of plasma, serum, or cell culture supernatant input
◦ Up to 10 unique analytes per well, and 175 samples in duplicate per 384-well plate
◦ Read out on high-content imagers, <20min scan/plate
● Consistent: ◦ <15% intra-plate CVs
◦ 70-130% sample recovery
◦ Choose from over 800 available antibody pairs ◦ Recombinant pairs, therefore no lot-to-lot variability
Assay Workflow:
FirePlex®-HT quantifies up to 10 protein analytes per well from low sample inputs. The two-step workflow and no-wash, 384-well plate assay format limit hands-on time and are amenable to automation, thus making FirePlex®-HT ideally suited for high-throughput screening studies. Assay readout is conducted on high-content imagers and data analysis is performed with the integrated, free-of-charge FirePlex® Analysis Workbench software, thus bypassing the need for dedicated instrumentation or expensive software licences.
Figure 1. To capture analytes onto FirePlex particles, biological samples are added to a 384-well imaging plate and incubated with the FirePlex-HT Reagent Mix overnight. Subsequently, an Imaging Dye is added and plates are scanned on high-content imagers (refer to Table 1 for a list of currently validated high-content imagers). The FirePlex Analysis Workbench software generates standard curves and quantifies analytes of interest directly from image files.
Image AnalyzeIncubate Samples + Particles
+
FirePlex® Reagent Mix
Biologicalsamples
1. Shakeovernight
High-content imager
Data analysis with FirePlex® Analysis
Workbench Software
Analytes captured onto fluorescently labelled particles
2. AddImaging Dye
3. CollectData
Key Assay Features:
We’d love your input on FirePlex-HT Immunoassays! Please contact [email protected]:• Provide suggestions for analytes of interest • Inquire about custom panel design • Become an early adopter
Copyright © 2018 Abcam, All rights reserved. FirePlex® is a registered trade mark in the United States and is an unregistered trade mark elsewhere.
Figure 2. A-B. Varying levels of green and yellow fluorescent dyes are used for particle identity barcoding, and a red fluorescent dye is used for protein analyte quantification. Three images are captured per well, with approximately 20 particles analyzed per analyte using the FirePlex Analysis Workbench software. C. Representative graph demonstrating red fluorescent intensity levels automatically output from scanned wells. D-F. Representative plots of an eight-point standard curve analysis for CXCL1 (D), IFNγ (E), and IL-1β (F) scanned on Molecular Devices ImageXpress Micro, PerkinElmer Opera Phenix, TTP Labtech mirrorball, and Yokogawa Cell Voyager 7000 high-content imagers and analyzed with the FirePlex Analysis Workbench software.
CXCL1 IFNγ IL-1β
A B C
D E F
Data Collection with High-Content Imagers
Validated High-content Imagers Available Analytes
Tables 1 and 2. (Table1) List of high-content imaging instruments currently validated for use with FirePlex-HT, and required specifications. (Table 2) Human analytes and corresponding sensitivities available with FirePlex-HT.
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