n Budget constraints n Limited benchspace n Interest in running magnetic n User-friendly data management n Benefits resulting from a
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1 Acute Phase Response Cancer Cardiovascular Disease Cytokines, Chemokines, Growth Factors Diabetes Gene Expression Genotyping Immunoglobulin Isotyping MicroRNA Cell Signaling Toxicology Bio-Plex MAGPIX Bio-Plex 2 and Bio-Plex 3D Systems Comparable Performance Across the Board Dedicated Magnetic advantage Practical, Versatile, and High-Throughput Options n Time savings n Cost effectiveness n Flexible multiplexing n Sample saving Bio-Rad offers a comprehensive selection of array readers for multiplex applications to satisfy a variety of research needs and budgets. These readers include the Bio-Plex MAGPIX system with practical multiplexing functionality, the versatile Bio-Plex 2 system, and the Bio-Plex 3D system with high-throughput capacity. Assay performance based on sample concentration determinations, working assay ranges, limit of detection (LOD), and inter-assay %CV is similar on the Bio-Plex MAGPIX, Bio-Plex 2, and Bio-Plex 3D systems. Bio-Plex Pro assays can be run on all three systems. Bio-Plex MAGPIX System Practical Multiplexing The Bio-Plex MAGPIX system is appropriate for labs that have n Budget constraints n Limited benchspace n Interest in running magnetic bead based immunoassays Bio-Plex 2 System Unparalleled Versatility The Bio-Plex 2 system is suitable for labs that value n User-friendly data management n Benefits resulting from a single acquisition and analysis software package n Running magnetic and nonmagnetic assays Bio-Plex 3D System High-Throughput Capacity The Bio-Plex 3D system is optimal for labs with n Fewer budget restrictions n High-throughput needs n Robotics-interfacing requirements
2 Bio-Plex Pro human 27-plex group I cytokine assays were tested on the Bio-Plex MAGPIX, Bio-Plex 2, and Bio-Plex 3D systems using 96-well plates. The performance of the Bio-Plex assays was compared based on sensitivity, assay range, precision, accuracy, and sample concentrations. The standards, blanks, and serum samples were run in duplicates using the low PMT setting on Bio-Plex 2 and Bio-Plex 3D systems. Comparable performance was observed from all three systems. Standard Curves from the Bio-Plex MAGPIX, Bio-Plex 2, and Bio-Plex 3D Systems Have Similar Shapes and Slopes Representative standard curves from human 27-plex assays were generated for data acquired from the three instruments (Figure 1). The standard curves have similar shapes and slopes. However, mean fluorescence intensity (MFI) values obtained on the Bio-Plex 3D system were higher than those of the Bio-Plex 2 system. A high PMT setting on the Bio-Plex 2 system generated curves similar to those of the Bio-Plex 3D System. These higher MFI values do not result in higher sensitivity. 1,, 1,, 1,, 1, 1, 1, 1, 1, 1, MFI 1, 1, 1, , 1, 1, , 1, 1, , 1, 1, 1,, 1,, IP-1 1,, 1, 1, 1, 1, 1, 1, MFI 1, 1, 1, , 1, 1, , 1, 1, , 1, 1, Fig. 1. Standard curve comparison of human 27-plex assays using the Bio-Plex MAGPIX (n), Bio-Plex 2 (n), and Bio-Plex 3D (n) systems. 212 Bio-Rad Laboratories, Inc. Bulletin 66
3 The Bio-Plex MAGPIX, Bio-Plex 2, and Bio-Plex 3D Systems Report Comparable LOD Values LOD is the concentration corresponding to the minimum MFI value that can be reliably differentiated from background and is defined as two standard deviations of the appropriate blank values. This translates into 99% certainty that any signal measured at this MFI is the result of the presence of the analyte. The LOD of human 27-plex assays tested are shown here as mean values from three independent assays (Figure 2). Results demonstrate that the sensitivity of the majority of analytes is comparable on all three instrument platforms IL-1 IL-12p7 IP-1 Fig. 2. LOD of Bio-Plex Pro human 27-plex group I cytokines using the Bio-Plex MAGPIX (n), Bio-Plex 2 (n), and Bio-Plex 3D (n) systems. The Bio-Plex 3D System Demonstrates Higher Background and Sample MFI Than Other Platforms The background signal assessed using a human 27-plex assay was similar on the Bio-Plex MAGPIX and Bio-Plex 2 systems, but was higher on the Bio-Plex 3D system (Figure 3). This corresponds to the proportionate increase in MFI at all points of the standard curve on the Bio-Plex 3D system MFI 1 5 IL-1 IL-12p7 IP-1 Fig. 3. Background signal obtained with human 27-plex assays using the Bio-Plex MAGPIX (n), Bio-Plex 2 (n), and Bio-Plex 3D (n) systems. Similar Inter-Assay Precision on All Three Instruments The inter-assay precision calculated using observed concentrations of spiked control from three independent experiments was comparable on all three instrument types for most of the analytes tested (Figure 4). Spiked controls were created by adding recombinant analytes to human serum %CV IL-1 IL-12p7 IP-1 Fig. 4. Inter-assay precision of human 27-plex assays using the Bio-Plex MAGPIX (n), Bio-Plex 2 (n), and Bio-Plex 3D (n) systems. 212 Bio-Rad Laboratories, Inc. Bulletin 66
4 Similar Working Assay Range The working assay range is defined as the standard curve range where the analytes meet the recovery of 7 13% and intra-assay precision of 2% and is bound by the lower limit of quantification (LLOQ) and upper limit of quantification (ULOQ). Assay ranges for most of the analytes were similar on all three instruments, with a few minor fluctuations in LLOQ (Figure 5). A B 12, IL-1 IL-12p7 IP-1 1, 8, 6, 4, 2, IL-1 IL-12p7 Fig. 5. Lower limit of quantification (A) and upper limit of quantification (B) obtained with the Bio-Plex MAGPIX (n), Bio-Plex 2 (n), and Bio-Plex 3D (n) systems. Similar Sample Concentrations on All Three Instruments Six serum samples were evaluated for concentration on the three instruments. Figure 6 depicts the concentrations of samples analyzed using the Bio-Plex MAGPIX, Bio-Plex 2, and Bio-Plex 3D systems for representative analytes from human 27-plex assays. Concentration values were similar for all samples tested with Bio-Plex Pro human 27-plex group I cytokines. The error bars represent standard deviations from three independent experiments. IP ,6 25 IL-12p7 25 1, Sample ID Sample ID Sample ID Fig. 6. Sample concentrations acquired with the Bio-Plex MAGPIX (n), Bio-Plex 2 (n), and Bio-Plex 3D (n) systems. 212 Bio-Rad Laboratories, Inc. Bulletin 66
5 Table 1. Instrument and assay compatibility. Assay Bio-Plex MAGPIX System Bio-Plex 2 System Bio-Plex 3D System Magnetic bead based assays (6.5 µm bead) Bio-Plex Pro human cancer biomarker panel assays Bio-Plex Pro human Th17 assays Bio-Plex Pro mouse Th17 assays Bio-Plex Pro cell signaling assays Bio-Plex Pro human cytokine assays* Bio-Plex Pro mouse cytokine assays Bio-Plex Pro rat cytokine assays Bio-Plex Pro human diabetes assays Bio-Plex Pro mouse diabetes assays Bio-Plex Pro rat diabetes assays Bio-Plex Pro TGF-b assays Nonmagnetic bead based assays Bio-Plex signal transduction assays Magnetic fixed panel assays (8. µm bead) Bio-Plex Pro acute phase assay Bio-Plex Precision Pro human cytokine assay Bio-Plex Pro isotyping assay Bio-Plex Pro angiogenesis assay * Except Bio-Plex Precision Pro human cytokine assay (catalog #171-A11P). Table 2. Distinct features of the three instrument platforms. Bio-Plex MAGPIX System Bio-Plex 2 System Bio-Plex 3D System Software xponent (acquisition) Bio-Plex Manager xponent (acquisition) Bio-Plex Manager (analysis) (acquisition and analysis) Bio-Plex Manager xponent (optional) (analysis) Optics LED/CCD camera Lasers/APDs/PMTs Lasers/APDs/PMTs Multiplex capacity Bead compatibility MagPlex C MagPlex C MagPlex C MicroPlex MicroPlex LumAvidin LumAvidin xtag xtag Read time ~6 min/96-well plate ~4 min/96-well plate ~2 min/96-well plate ~75 min/384-well plate Applications Low- to mid-throughput Mid-throughput High-throughput Plate format 96-well 96-well 96-well/384-well Dynamic range 3.5 logs 3.5 logs 4.5 logs Consistent Performance Across All Platforms The assay performance based on sample concentration determinations, working ranges, LOD, and inter-assay %CV is similar on Bio-Plex MAGPIX, Bio-Plex 2, and Bio-Plex 3D systems. Bio-Plex Pro assays can be run on all three systems. 212 Bio-Rad Laboratories, Inc. Bulletin 66
6 Ordering Information Catalog # Description Instruments Bio-Plex MAGPIX Multiplex Reader with Bio-Plex Manager Software, includes Bio-Plex MAGPIX instrument, PC with xponent acquisition software installed, Bio-Plex Manager desktop license, calibration kit, verification kit, 2 drive fluid cartridges, 2 waste containers Bio-Plex 2 System, 1 24 V, includes array reader, microplate platform, Bio-Plex Manager software with workstation license, PC, monitor, calibration kit, validation kit 4., MCV plate IV, Bio-Plex reservoir, 2 L sheath fluid, spare sample needle Bio-Plex 2 System with HTF, same as with high-throughput fluidics (HTF) Bio-Plex 3D Bio-Plex 3D System, includes Bio-Plex 3D suspension array system, xponent acquisition software, PC, calibration reagents, verification reagents, Bio-Plex Manager software, desktop license Kits and Reagents Bio-Plex MAGPIX Calibration Kit, calibration testing kit for Bio-Plex MAGPIX instrument, includes 5 ml calibrator microspheres, twenty-five 8-well strips, CD; 25 applications Bio-Plex MAGPIX Verification Kit, includes MAGPIX verifier, 5 ml microspheres, MAGPIX fluidics 1, MAGPIX fluidics 2, MAGPIX performance verification kit CD, twenty-five 8-well strips; 25 applications Bio-Plex MAGPIX Drive Fluid, pkg of 4, 7 ml drive fluid, for use with Bio-Plex MAGPIX instrument Catalog # Description Software Bio-Plex Data Pro Software with Bio-Plex Manager Software, Bio-Plex Data Pro software (5 seats), for multi-experiment analysis and advanced data visualization, and Bio-Plex Manager software (5 seats), for instrument data evaluation and optimization. CDs and security HASP key included Bio-Plex Data Pro Software, (5 seats), for multi-experiment analysis and advanced data visualization 171-STND1 Bio-Plex Manager Software, includes 1 user desktop license, to analyze Bio-Plex data and generate protocols, does not operate the instrument Luminex, MAGPIX, MagPlex, MicroPlex, LumAvidin, xmap, xponent, and xtag are trademarks of Luminex Corporation. HASP is a trademark of Aladdin Knowledge Systems, Ltd. The Bio-Plex suspension array system includes fluorescently labeled microspheres and instrumentation licensed to Bio-Rad Laboratories, Inc. by the Luminex Corporation. CST antibodies exclusively developed and validated for Bio-Plex phosphoprotein and total target assays. Accessories Bio-Plex MAGPIX Replacement Waste Fluid Container, pkg of 1, 85 ml container, holds waste fluid, for use with Bio-Plex MAGPIX instrument Bio-Plex MAGPIX Sample Probe Needle, pkg of 1, needle Bio-Plex MAGPIX Sample Probe Height Adjustment Kit, sample probe height adjustment kit Bio-Plex MAGPIX 96-Well Plate Heater Block, pkg of 1, heater block, fits one 96-well plate, for use with Bio-Plex MAGPIX instrument Bio-Plex Handheld Magnetic Washer, includes magnetic washer and adjustment hex tools for use in manual wash steps for all Bio-Plex magnetic assays Bio-Plex Pro Wash Station, includes magnetic plate carrier, waste bottle, 2 buffer bottles Bio-Plex Pro II Wash Station, includes magnetic plate carrier, vacuum manifold plate carrier, waste bottle, 2 buffer bottles Bio-Rad Laboratories, Inc. Life Science Group Web site USA Australia Austria Belgium Brazil Canada China Czech Republic Denmark Finland France Germany Greece Hong Kong Hungary India Israel Italy Japan Korea Mexico The Netherlands New Zealand Norway Poland Portugal Russia Singapore South Africa Spain Sweden Switzerland Taiwan Thailand United Kingdom Bulletin 66 Rev A US/EG Sig 1211
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