Mouse IgG Type and Titer Assay

Key Benefits

• Speed: Sample to actionable data workflow time—assay set up, incubation, read—under 100 minutes per 384 well plate. Real time analysis and visualization from one powerful software package: ForeCyt.
• Results: Five endpoints from a single multiplexed, no-wash/no-dilution assay: IgG isotype identification, IgG quantity per isotype, total IgG secretion level, cell viability, and cell counts.
• Simplicity: Single Platform Assay Development and Data Acquisition eliminates the time and confusion of acquiring, analyzing and correlating data from multiple assays on multiple platforms.
• Precision: Isotyping facilitates specific primer design for superior PCR-mediated gene cloning. Cell quality criteria determine the best clones for RNA isolation.

Kit Description

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:

• 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.

Workflow

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

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 Cell 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.