High-Throughput Screening (HTS) Cytometry Reagents for Antibody Characterization

The only solution that correlates IgG quantitation per cell with cell health and growth readouts in a single, cost-effective competition assay using IgG-capture beads, IgG fluorescent competing antibody, and a cell viability stain.

Clone productivity ranking with more relevant data—Balancing IgG concentration, cell density, IgG quantitation per cell, and cell viability are crucial to selecting the optimal clones.
 

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The Mouse IgG Type and Titer kit is a multiplexed, competition formatted assay to identify and quantify mouse IgG1, IgG2a, IgG2b, and IgG3.  It is a no wash, no sample dilution assay that measures both cells and beads and provides these significant advantages over traditional methods:
 

Read the Mouse IgG Type and Titer Kit Application Note

• IgG isotype identification — Guides the decision-making process by identifying monoclonal wells, thus reducing the number of cell sub-cloning steps while facilitating isotype-specific DNA primer sets rather than degenerate primers for cloning or sequencing of antibody variable regions.
• Precise total and isotype specific IgG quantification — Downstream confirmatory or functional assays benefit from normalizing the IgG concentration for all clones.
• Simultaneous measurement of cell count and cell viability — Identifies the cells healthy enough for the RNA extraction needed for PCR-mediated antibody cloning.

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Streamlined workflow means experiment set-up and data acquisition for this multiplex assay takes about 100 minutes—and that includes incubation.

iQue^® Mouse IgG Type and Titer Workflow. Samples from hybridoma or B-cell cloning plates are directly transferred to the assay plate without an intermediate dilution step. Each assay well is seeded with four different types of capture beads, each with a specific affinity for a single IgG isotype. The IgG isotype in the sample will compete with the same FITC-labeled IgG isotype to bind to the capture bead. The IgG isotype quantity will be calculated from the isotype-specific standard curve.  If cells are included in the assay well, a membrane integrity dye in the detection reagent will stain the membrane-compromised dead cells by DNA intercalation. After the addition of the detection reagent mixture and the capture beads to the samples and 60 minute incubation, the samples are acquired on the iQue Screener PLUS.