Introducing FirePlex-HT


High-throughput features, without compromising on high-performance multiplex analysis

FirePlex®-HT immunoassays offer the high-standards of performance and multiplexing you expect from our original FirePlex immunoassays, now in a high-throughput, automation friendly format.  

Ideal for discovery, screening and profiling projects, Fire-Plex-HT offers you a fast, easy and cost-effective tool to analyze thousands of samples rapidly. Reach your R&D goals faster, and accelerate your path to success with FirePlex-HT immunoassays.



Drug development applications

FirePlex-HT can be applied at all stages of drug development to inform critical decision points. By providing a new, more effective and affordable biomarker analysis workflow, FirePlex-HT enables data-driven R&D to guide the most effective strategies and minimize the risk of late stage failures.

  • Discovery projects screening large compound libraries
  • Lead optimization, safety assessment and toxicity screening in large sample sets
  • Clinical screening for disease identification, patient stratification or therapy response in large patient cohorts


​Accelerate your research

FirePlex-HT uses our custom FirePlex particle technology and our highly validated antibody pairs to deliver rapid, flexible, and sensitive high-throughput multiplex screening without the need for proprietary or closed system instruments. 

NEW automation-friendly features to accelerate work-flow:

  • 384-well plate assay format
  • No wash, two-step workflow
  • Read out on high-content imagers, < 20 min scan/plate

Analyze up to 10 analytes per sample, direct from plasma, serum or cell culture supernatant — no pre-processing required

The assay workflow

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. The FirePlex Analysis Workbench software generates standard curves and quantifies analytes of interest directly from image files.


Scale up without sacrificing performace

assay performance or data quality.
1-100 pg/mL sensitivity (analyte dependent)
2.5-3 logs dynamic range
only 6.5 µL biofluid input (plasma, serum or cell culture supernatant)
<15% intra-plate CVs
recombinant antibody pairs eliminate lot-to-lot variability
70-130% sample recovery
reread plates up to a week later, as many times as needed

With FirePlex-HT there is no trade-off between quantity and quality. Achieve high-throughput without sacrificing assay performance or data quality.

  • 1-100 pg/mL sensitivity (analyte dependent)
  • 2.5-3 logs dynamic range
  • 6.5 µL biofluid input (plasma, serum or cell culture supernatant)
  • <15% intra-plate CVs
  • Recombinant antibody pairs eliminate lot-to-lot variability
  • 70-130% sample recovery
  • Reread plates up to a week later, as many times as needed


Table 1. Human analytes and corresponding sensitivities available with FirePlex-HT.


Analyte

Sensitivity (pg/mL)

Analyte

Sensitivity (pg/mL)

BCA1

16.19

IL-4

16.39

CXCL2

6.77

IL-5

2.69

Eotaxin

34.95

IL-6

5.14

Fractalkine

10.81

IL-7

22.33

GROα

0.35

IL-8

7.84

IL-1β

2.06

IL-9

22.69

IL-2

6.88

IL-10

58.03

IL-13

20.53

IFNα1

13.10

IL-12 p70

11.58

IFNα2

53.10

IL-17A

1.27

IFNβ

79.09

IL-1Ra

5.58

IFNγ

64.80

IP-10

15.71

RANTES

0.58

MCP1

1.50

TARC

115.17

MDC

13.47

TNFα

36.60


Flexibility in analytes

Choose from 

  • Over 800 available antibody pairs
  • Top 40 key cytokines and subsets of critical organ toxicity markers 
  • ~70 human analytes and ~30 mouse analytes
  • Pre-designed focus panel or custom panel options


Fast and automation friendly

  • 6.25 µL of plasma, serum or cell culture supernatant input
  • No wash, two-step workflow
  • Readout on high-content imager with <20 min scan/plate
  • Analyze up to 175 samples in duplicate per 384-well plate


Pre-designed and customizable panels

Pre-designed 10-plex and 6-plex focus panels offer ready-to-go analysis of key human cytokine combination.

Review product-specific certificates of analysis (CoA) here

These human cytokine kits contain the FirePlex particle mix and all necessary reagents to quantify the indicated human cytokines. The kits are supplied with the required volume of reagents to assay 1 x 384-well plate

Customize your own your own panels to fit your specific research needs. Flexible analyte selection allows you to choose from:

  • over 800 available antibody pairs
  • ~70 human analytes and ~30 mouse analytes
  • top 40 key cytokines and subsets of critical organ toxicity markers
  • “pick-and-mix” panels based on biological relevance for quick and economical panel assembly


Simple data collection and analysis

Scan data quickly and easily on high content imagers, in less than 20 minutes per plate. FirePlex-HT assays are compatible with machines from various prominent manufacturers including Molecular Devices, TTP Labtech, Perkin Elmer, GE, and Yokagawa. To facilitate instrument set-up, we also offer a FirePlex-HT Imager Setup Kit (ab229130) .

FirePlex-HT uses levels of green and yellow fluorescent dyes to identify individual particles and a red fluorescent dye for protein quantification. This allows for comparative analysis between a range of high content imagers. (Find out more about FirePlex particle technology)

With integrated and free-of-charge FirePlex Analysis Workbench software, complex data analysis is made simple.  The sophisticated, yet easy-to-use software decodes data files, performs standard curve analysis, quantitates analytes, and generates publication-quality plots and graphs which can be easily exported for manuscript preparation. ​​

With FirePlex-HT there is no trade-off between quantity and quality. Achieve high-throughput without sacrificing assay performance or data quality.
1-100 pg/mL sensitivity (analyte dependent)
2.5-3 logs dynamic range
only 6.5 µL biofluid input (plasma, serum or cell culture supernatant)
<15% intra-plate CVs
recombinant antibody pairs eliminate lot-to-lot variability
70-130% sample recovery
reread plates up to a week later,

Figure 2. A–B: Fluorescent particle output. Three images, on in each channel (Ai–Aiii) are captured per well with approximately 20 particles analyzed per analyte using the FirePlex Analysis Workbench software. C–E: Eight-point standard curve analysis for CXCL1 (C), IFNγ (D), and IL-1β (E) 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.


Automation

FirePlex-HT is ideally suited to high-throughput automated workflows. 

 A: Example of an automated workflow for the FirePlex-HT Immunoassay. The Reaction Mix is dispensed into the Assay Plate using a ThermoFisher Multidrop Combi (Step 1), followed by transfer of biological samples (Step 2) and Assay Diluent (Step 3) into the Assay Plate using a liquid handler with a 384 channel pipette head. Samples are incubated overnight at room temperature, followed by addition of the Imaging Dye into the Assay Plate using a Multidrop Combi (Step 4) and image acquisition with a high-content imager (Step 5). B: Representative dataset from a FirePlex-HT immunoassay experiment run with an automated workflow. Data are interpolated concentrations of CXCL1 in untreated and treated cell culture supernatants.


Compatible high content imagers

Readout of FirePlex-HT Immunoassays is compatible with various instrumentation platforms, including those manufactured by Molecular Devices, TTP Labtech, Perkin Elmer, GE, and Yokagawa.


Field of view (on 384-well plate)DetectionColorExcitation (nm)Emission (nm)
Entire single well16-bit  sCMOS  Camera(s)Green440-500500-535
Entire single well16-bit  sCMOS  Camera(s)Yellow440-500570-630
Entire single well16-bit  sCMOS  Camera(s)Red490-560650-760

We offer a FirePlex-HT Imager Setup Kit (ab229130) desgined to identify properly resolving high-conetnt imaging instruments, which can be validated for use with FirePlex-HT immunoassays.


Sample profiling services

Send us your samples for profiling and let us do the work. Our team of FirePlex® experts offers full support, from generating hypotheses to delivering results. With experience accumulated over testing 10,000 customer samples, you can confidently entrust your research to our skilled scientists. Results, along with publication-ready charts, tables and raw data files, are typically delivered within approximately 15 business days.


FirePlex-HT flyer

FirePlex-HT poster


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