Blood plasma was analyzed using the Milliplex Map Human Magnetic Bead Panel for cytokines and chemokines (HCYTMAG-60 K-PX41) according to the manufacturer’s protocol on a Bio-Plex 3D instrument (Bio-Rad) as done previously33 (link). The ELISA assay details, and analyte classification are given in Appendix Table 1 .
Bio plex 3d instrument
Manufactured by Bio-Rad
Sourced in United States
The Bio-Plex 3D instrument is a high-performance multiplex assay system that enables simultaneous detection and quantification of multiple analytes in a single sample. It utilizes magnetic beads with unique spectral addresses to provide precise and sensitive measurements. The instrument features advanced optics and fluidics for efficient sample processing and data acquisition.
Automatically generated - may contain errors
5 protocols using bio plex 3d instrument
1
Multiplex Cytokine Profiling of Blood Plasma
2
Quantifying Alzheimer's Biomarkers in CSF
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CSF Aβ42, Aβ40, total tau (tTau), and phosphorylated threonine 181 tau (pTau) were measured using MILLIPLEX® MAP Human Amyloid Beta Tau Magnetic Bead Panel (Luminex xMAP) run on the Bio-Plex® 3D instrument (Bio-Rad Laboratories, Hercules, CA) according to manufacturer’s instructions. The kit allows simultaneous quantification of Aβ40, Aβ42, tTau, and pTau.
3
Cytokine Analysis of Distal Colon Tissue
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Distal colon tissue was washed in cold PBS and directly frozen in 500 μL of PBS containing 0.5% BSA, 0.5% Tergitol and protease inhibitor (Sigma-Aldrich). Tissue was lysed using a steel bead and a Qiagen tissue lyser (10 min, full speed) and centrifuged (20 min, 5000 g, 4°C). Supernatant was used undiluted for bioplex assay (Biorad). Concentration of the following cytokines was analyzed using the standard manufacturer’s protocol: GM-CSF, KC, IFN-g, IL-1b, IL-6, IL-10, IL-7a, IL-22, IL-18, IL-23p19, IL-33, MCP-1 (CCL2), and MIG (CXCL9). Samples were acquired using the Bio-Plex 3D instrument (Biorad) and the xPONENT Software (Luminex). Data was analyzed using the Bioplex Manager 6.1 (Biorad).
4
Quantifying Antibody Profiles in Typhoid IS
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The total concentration of antibody within 16/138 WHO typhoid IS was quantified using the Bio-Plex Pro Human Isotyping Assay kit per manufacturer's instructions (26 (link)). Briefly, anti-human IgA, IgM, IgG1, IgG2, IgG3, and IgG4-conjugated magnetic microspheres were incubated with titrated 16/138 WHO typhoid IS serum followed by primary and secondary detection. Fluorescence intensity (FI) was collected using the Bio-Plex 3D instrument (Bio-Rad, USA). The isotype and subclass components of the international serum standard, 16/138 typhoid IS, were determined with this method (5.69 mg/ml IgG1; 6.84 IgG2; 0.57 IgG3; 0.58 IgG4; 1.38 IgM; 0.96 IgA mean mg/ml, of 6 technical replicate measurements). These concentration units were utilized to calculate antibody subclass concentration in μg/ml equivalents of 16/138 in vaccinee sera. The 16/138 Typhoid IS IgA equivalents are noted for the non-IgG depleted standard.
5
Cytokine and Immunoglobulin Profiling from Frozen Stool
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Frozen stool samples were thawed, and approximately 4 mg of each sample were dissolved in 200 μl of phosphate buffered saline. The supernatant obtained by centrifugation at 16,000 g for 15 min was then analyzed using the Milliplex Map Human Magnetic Bead Panels for cytokines and chemokines (HCYTMAG-60K-PX41) and immunoglobulins (HGAMMAG-301K-06) according to the manufacturer’s protocol on a Bio-Plex 3D instrument (Bio-Rad). The ELISA assay details and analyte classification are given in Supplementary Table 1 . To avoid the bias of inter-plate variation, paired samples were examined on the same ELISA plate.
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