Metabolism Cancer Cardiovascular Disease Cytokines, Chemokines, Growth Factors Neurology Diabetes Infectious Disease ™ Inflammation Bio-Plex Pro Signal Transduction Sepsis Human 10-Plex Assay

Apolipoprotein A1 / / / Apolipoprotein C1 / Apolipoprotein C3 MAGNETIC SEPARATION ENABLED Apolipoprotein D / / / Apolipoprotein J / C-Reactive

n All-in-one High-Performance Multiplex Assay Features premixed kit Immunoassays for Research This panel is offered in a convenient, n Optimized The Bio-Plex Pro Human Apolipoprotein Assay all-in-one, 10-plex kit format that includes for lot-to-lot Panel is a sensitive, magnetic bead–based magnetic capture beads, detection antibodies, reproducibility multiplex assay that allows you to accurately vial of standards, two-level controls, diluents, n Two-level measure nine and C-reactive buffers, streptavidin-PE, flat bottom plate, and quality controls protein (CRP) in diverse matrices, including plate seals for the detection of nine human n Magnetic workflow serum and plasma. Multiplex capabilities apolipoproteins and CRP (Table 1). allow you to rapidly quantitate multiple ■■ Manufactured in accordance with apolipoproteins in a single microplate well GMP guidelines in just 4 hours, using only 10 µl of sample. ■■ Lot-to-lot correlation specification of Validated to rigorous analytical standards and R2 ≥ 0.9 for consistent, reproducible results designed for lot-to-lot consistency, this panel ■■ Full multiplate validation documentation is ideal for investigating biomarker profiles for each kit lot associated with the following research areas: ■■ Two-level quality controls with lot-specific ranges ■■ Cardiovascular disease ■■ Assay quick guide to get you started ■■ Inflammation right away ■■ Diabetes ■■ Fastest available assay protocols and ■■ Neurological disease a convenient all-in-one kit format ■■ Sepsis ■■ Compatible with the Bio-Plex® 200, Bio-Plex ■■ Cancer 3D Suspension Array Systems, and the Bio-Plex® MAGPIX™ Multiplex Reader ■■ Magnetic beads for simplified plate processing

Table 1. Analytes detected by the Human Apolipoprotein Panel. Apo A1 Apo C3 Apo H Apo A2 Apo D Apo J Apo B Apo E CRP (C-reactive protein) Apo C1 Table 2. Representative performance characteristics. above the background signal (limit ofSensitivity detection, LOD) range working assay the within (recovery) Accuracy Precision (ULOQ) of quantitation limit upper the and (LLOQ) of quantitation limit lower by the defined are range Assay working range Table in 2. shown as performance, of assay indicative are parameters following The Assay Performance Definitions comparing the standard curves of assay controls to biological samples. to biological controls of assay curves standard the comparing by demonstrated as assay of the regions quantifiable the within fall samples biological that to ensure optimized and developed are Assays Pro Bio-Plex research. in to useful be order in of expression range biological the encompass should range working assay The Working Range ** * C-reactive protein Apolipoprotein J Apolipoprotein H Apolipoprotein E Apolipoprotein D Apolipoprotein C3 Apolipoprotein C1 Apolipoprotein B Apolipoprotein A2 Fluorescence intensity Analyte extracting the concentration from the standard curve. All values in the table represent the mean from at least three separate assay plates. assay separate three least at from mean the represent table the in values All curve. standard the and from diluent standard concentration of the (MFI) extracting intensity fluorescence median background to the deviations standard two adding by determined detection of limit CV). (≤30% the is LOD precise and (80–120% recovery) accurate both are measurements where quantitation of limits upper and lower the are ULOQ and LLOQ 20,000 10,000 0 – the coefficient of variation (%CV) at concentrations within the assay working range working assay the within concentrations at (%CV) of variation coefficient –the ULOQ (61.127)ULOQ

0.01 S8 (0.03) S8 Clusterin/Apo J – percentage of the observed concentration relative to the expected concentration of a known amount of analyte of analyte amount of aknown concentration to expected the relative concentration observed of the –percentage Alternative – the range of concentrations within which the assay is precise and accurate. Boundaries of the assay working working assay of the Boundaries accurate. and precise is assay the which within of concentrations range –the S7 (0.10) S7 Apo C3 Apo Apo C1 Apo Names Apo A2 Apo Apo A1 Apo Apo H Apo Apo D Apo Apo B Apo Apo E Apo 0.10 CRP S6 (0.29) S6 Concentration, ng/ml Concentration, Apolipoprotein A1 S5 (0.86) S5 Region 1.00 – the concentration of analyte for which the fluorescence intensity signal is two standard deviations deviations standard two is signal intensity fluorescence the which for of analyte concentration –the Bead 48 38 36 44 39 22 26 78 75 12 S4 (2.61) S4 Assay Working Ranges, ng/ml Ranges, Working Assay 10.00 S3 (7.72) S3 LLOQ* 0.030 0.055 0.055 0.059 0.032 0.023 0.021 0.019 0.41 0.15 0.12 S2 (24.11) S2 LLOQ (0.032) LLOQ S1 (61.13) S1 100.00 360 360 210 170 36 36 70 70 ULOQ* 30 28 12 17 11

Fluorescence intensity 20,000 10,000 0 Assay Sensitivity, ng/ml ULOQ (22.935) 0.01 0.0082 0.0087 LOD** 0.045 0.027 0.078 0.016 0.013 0.012 0.22 S8 0.11 S7 (0.03) S7 S6 (0.09) S6 0.10 Concentration, ng/ml Concentration, Apolipoprotein A2 Intra-Assay %CVIntra-Assay S5 (0.28) S5 2 3 3 3 3 4 3 6 6 4 1.00 S4 (0.84) S4 Assay Precision S3 (2.55) S3 Inter-Assay %CV 10.00 S2 (7.67) S1 (22.94) S1 10 15 12 8 8 5 9 6 5 7 LLOQ (0.032) (0.032) LLOQ Fluorescence intensity Fluorescence intensity Fluorescence intensity 10,000 15,000 20,000 10,000 20,000 10,000 5,000 0 0 0

0.00 ULOQ (7.500) ULOQ (6.003) ULOQ (171.327) ULOQ S8 (0.01) S8 S8 S8 0.01 0.10 S7 (0.01) S7 0.01 S7 (0.27) S7 S7 (0.02) S7 S6 (0.03) S6 S6 (0.69) S6 S6 (0.08) S6 1.00 0.10 Concentration, ng/ml Concentration, Concentration, ng/ml Concentration, Concentration, ng/ml Concentration, Apolipoprotein C3 S5 (0.09) S5 Apolipoprotein E Apolipoprotein B 0.10 S5 (0.22) S5 S5 (2.19) S5 S4 (0.29) S4 10.00 S4 (0.29) S4 1.00 S4 (0.67) S4 S4 (7.04) S4 1.00 S3 (0.82) S3 S3 (2.00) S3 S3 (19.51) S3 S2 (2.50) S2 100.00 S2 (60.43) S2 S2 (6.00) 10.00 10.00 S1 (7.50) S1 LLOQ (0.012) LLOQ S1 (171.33) S1 LLOQ (0.008) (0.008) LLOQ LLOQ (0.271) (0.271) LLOQ

Fluorescence intensity Fluorescence intensity Fluorescence intensity 20,000 10,000 10,000 15,000 20,000 10,000 5,000 0 0 0 ULOQ (149.128)ULOQ ULOQ (8.324) ULOQ ULOQ (11.999) 0.01 S8 S8 S8 0.10 0.01 S7 (0.03) S7 S7 (0.20) S7 S7 (0.02) S7 S6 (0.64) S6 S6 (0.05) S6 S6 (0.11) S6 0.10 1.00 Concentration, ng/ml Concentration, Concentration, ng/ml Concentration, Concentration, ng/ml Concentration, 0.10 Apolipoprotein C1 Apolipoprotein D Apolipoprotein H S5 (1.78) S5 S5 (0.15) S5 S5 (0.33) S5 1.00 S4 (0.87) S4 S4 (0.45) S4 S4 (5.75) S4 10.00 1.00 S3 (16.30) S3 S3 (2.85) S3 S3 (1.33) S3 10.00 S2 (50.68) S2 S2 (8.32) S2 (4.00) S2 100.00 10.00 S1 (12.00) S1 S3 (149.13) S3 LLOQ (0.200) (0.200) LLOQ LLOQ (0.031) LLOQ LLOQ (0.017) LLOQ Apolipoprotein J CRP

ULOQ (99.701) S1 (99.70) ULOQ (6.999) S1 (7.0 0)

20,000 20,000

S2 (33.71) S2 (2.33) 10,000 10,000 Fluorescence intensity Fluorescence Fluorescence intensity Fluorescence S3 (10.77) S3 (0.77)

S4 (3.86) LLOQ (0.54) S4 (0.27) LLOQ (0.010) 0 0 S8 (0.05) S7 (0.13) S6 (0.40) S5 (1.240) S8 S7 (0.01) S6 (0.03) S5 (0.08)

0.01 0.10 1.00 10.00 0.00 0.01 0.10 1.00 10.00 Concentration, ng/ml Concentration, ng/ml

Fig. 1. Sample detection and working range. Standard curves with assay controls and serum samples were generated per the kit protocol. Standard points were prepared by serially diluting a reconstituted standard provided in each kit to generate an eight-point standard curve. The standard calibration curves plotted with standard points ( ), samples ( ), and controls ( ) illustrate the broad working range of the assays. Shown below are representative data for serum samples from healthy control groups and patients with cancer (prostate, lung, colorectal, and ovarian). n = 63.

Ordering Information Catalog # Description Catalog # Description Software 171001510 Bio-Plex Data Pro™ Software with Bio-Plex Manager™ Software, Human Apolipoprotein Assays Bio-Plex Data Pro Software (5 seats), for multi-experiment 12003081 Bio-Plex Pro Human Apolipoprotein Assay Panel, 10-plex, analysis and advanced data visualization, and Bio-Plex Manager 1 x 96-well, includes magnetic capture beads, detection antibodies, Software (5 seats), for instrument data evaluation and optimization. vial of standards, two-level controls, diluents, buffers, strepdavadin- CDs and security HASP key included PE, flat bottom plate, and plate seals for the detection of nine human 171001513 Bio-Plex Data Pro Software, (5 seats), for multi-experiment apolipoproteins and C-reactive protein analysis and advanced data visualization Wash Stations and Accessories 171STND01 Bio-Plex Manager Software, includes 1 user desktop 30034376 Bio-Plex Pro Wash Station, microplate wash station for license, for analysis of Bio-Plex data and generation of protocols, magnetic bead–based assays, includes magnetic plate carrier, does not operate the instrument waste bottle, 2 liquid bottles 171020100 Bio-Plex Handheld Magnetic Washer, includes magnetic Visit bio-rad.com/apolipoprotein for more information. washer and adjustment hex tools for use in manual wash steps for all Bio-Plex magnetic assays 171025001 Bio-Plex Pro Flat Bottom Plates, pkg of 40, 96-well plates, The Bio-Plex Suspension Array System includes fluorescently labeled for use with Bio-Plex Pro Wash Stations when using magnetic microspheres and instrumentation licensed to Bio-Rad Laboratories, Inc. bead–based assays by the Luminex Corporation. MAGPIX is a trademark of the Luminex Corporation.

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