Mice serum samples were collected 7 days after the last immunization and cytokine levels were measured. Mouse cytokine grp I panel 23-plex liquid-phase suspension microarray cytokine levels, including Th2/Th17 immune response (IL-4, IL-5, IL-6,IL-9, IL-10 and IL-17A) and Th1 immune response (IL-1α, IL-1β, IL-2, IL-3, IL-12p40, IL-12p70, IL-13, eotaxin, G-CSF, GM-CSF, IFN-γ, CXCR1, MCP, MIP-1α, MIP-1β, RANTES and TNF-α), were obtained using a Luminex 200™ system (Luminex Corporation, Netherlands) with a Bio-Plex Handheld Magnetic Washer (Bio Rad, USA). All data were processed using the Bio-Plex Manager software (Ver. 8.1, Bio Rad, USA).
Bio plex handheld magnetic washer
Manufactured by Bio-Rad
Sourced in United States
The Bio-Plex Handheld Magnetic Washer is a compact and portable device designed for use with Bio-Rad's suspension array systems. Its core function is to efficiently wash magnetic beads during immunoassay protocols.
Automatically generated - may contain errors
Lab products found in correlation
Bio plex pro mouse cytokine 23 plex assay, by Bio-Rad (1 mentions)
Bio plex manager software, by Bio-Rad (1 mentions)
Bio plex 200, by Bio-Rad (1 mentions)
45 plex human procartaplex panel 1, by Thermo Fisher Scientific (1 mentions)
Bio plex 200 system, by Bio-Rad (1 mentions)
Bio plex manager 6, by Bio-Rad (1 mentions)
Bio plex pro mouse cytokine 23 plex assay kit, by Bio-Rad (1 mentions)
Allegra x 22r centrifuge, by Beckman Coulter (1 mentions)
Oscillator, by Thermo Fisher Scientific (1 mentions)
Desk centrifuge 5418, by Eppendorf (1 mentions)
5 protocols using bio plex handheld magnetic washer
1
Cytokine Profiling of Immunized Mice
2
Cytokine Quantification in Murine Serum
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Serum cytokines at 12-weeks post-injury were quantified using the Bio-Plex Pro Mouse Cytokine 23-plex Assay (Bio-RAD, CA, United States) for IL-4, IL-6, IL10, IL-12(p40), G-CSF, IFN-γ, MIP-1α, MIP-1β Serum was diluted by 1:4, and 25 μL was loaded into each well. The plate was prepared as per the manufacturer’s instruction and washed using the Bio-Plex Handheld Magnetic Washer (Bio-RAD). The assays were run on the Bio-Plex 200 (Bio-RAD) and fluorescence intensity was collected. The calibration curve was analyzed using the Bio-Plex manager software (version 5.0, Bio-RAD).
3
Cytokine Profiling of 3D Cell Cultures
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The supernatant was collected and snap-frozen after completion of the 3D in vitro culture. The supernatants were then thawed and subjected to cytokine analysis using the Cytokine/Chemokine/Growth Factor Convenience 45-Plex Human ProcartaPlex™ Panel 1 (#EPXR450-12171-901, ThermoFisher, Waltham, MA, USA) following the manufacturer’s instructions. Wells were pooled and measured in duplicate. The samples were incubated with magnetic beads for 2 h. Plates were washed manually with the Bio-Plex® handheld magnetic washer (#171020100, BioRad, Hercules, CA, USA), then measured using a Bio-Plex® 200 System (#171000201, BioRad, Hercules, CA, USA).
4
Multiplex Cytokine Profiling in Mice
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A Bio-Plex Pro Mouse Cytokine 23-plex Assay kit (Bio-Rad Laboratories, Inc., Philadelphia, PA) was used for measuring cytokine content using a Bio-Plex 200 instrument (lab of Dr. Stanislaw Stepkowski, The University of Toledo, Toledo, OH) and Bio-Plex Manager 6.1 software (Bio-Rad Laboratories, Inc., Philadelphia, PA) was used for machine operation and data collection. A Bio-Plex handheld magnetic washer (Bio-Rad Laboratories, Inc., Philadelphia, PA) was used for all washing steps. The protocol accompanying the kit was followed. The following cytokines were measured: Eotaxin, G-CSF, GM-CSF, IFN-γ: Interleukins (IL’s); IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12(p40), IL-12(p70), IL-13, IL-17A, KC (keratinocyte), MCP-1/MCAF (monocyte chemotactic and activating factor), MIP-1α, MIP-1β (macrophage inflammatory protein), RANTES (regulated on activation, normal T cell expressed and secreted) and TNF-α (tumor necrosis factor alpha).
5
Multiplexed Immunoassay Protocol
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MILLIPLEX Multiplexing Assays (Merck & Co., Inc., Darmstadt, Germany); Millipore-Q Academic (Merck & Co., Inc., Darmstadt, Germany); Allegra x-22R Centrifuge (Beckman coulter, Inc., Brea, CA, USA); SpectraMax I3X Enzyme marker (Molecular Devices Instruments Ltd., San Jose, CA, USA); Desk centrifuge 5418 (Eppendorf, Inc., Hamburg, Germany); Vortex mixing device ORTEx Genius (IKA, Inc., Staufen, Germany); Digital ultrasonic cleaner (Kunshan Ultrasonic Instruments Co., Ltd., Kunshan, China); Bio-Plex Handheld Magnetic Washer (Bio-Rad Laboratories, Inc., Berkeley, CA, USA); Oscillator (Thermo Scientific, Co., Ltd., Waltham, MA, USA); Magnetic Shelf DvnaMag-2 (Thermo Scientific, Co., Ltd., Waltham, MA, USA); Biochemical incubator SPX-150B (Shanghai Pudong Rongfeng Scientific Instrument Ltd., Shanghai, China); 7600 Series Automatic Analyzer (HITACH Diagnostic Product Shanghai Ltd., Shanghai, China); Roche E601 Immunoluminescence instrument (Roche, Inc., Basel, Switzerland).
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