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ELISA Overview
Rapid Isotyping Arrays
1-hour Isotype Screen for Hybridoma Supernatants or Mouse Body Fluids
The growth and widespread use of mouse monoclonal antibody technology has created a need for a rapid, simple, and accurate means of determining immunoglobulin class & sub-class. Detection of specific Igisotypes is also a powerful tool in the study of immunoglobulin deficiency disorders, allergies, autoimmune diseases, malignancies, gastrointestinal disorders or repeated bacterial infections.
The Raybio® Rapid Mouse IgIsotyping Array is a glass chip-based multiplex assay which detects the 8 mouse immunoglobulin sub-classes (IgG1, IgG2a, IgG2b, IgG3, IgA, IgD, IgE, and IgM) plus the two light chain types (λ and κ). The array uses the sandwich ELISA-based format. However, unlike traditional ELISA which requires multiple steps and washes, the Rapid Isotyping Array features a streamlined, single incubation procedure that requires only 60 minutes of processing time. Moreover, up to 16 samples can be assayed simultaneously on one glass chip.
How It Works
First, samples are mixed with the Cy3 equivalent dye-labeled detection antibody, and then applied to the array slide. After a wash step, the slides are then dried and scanned with a compatible laser scanner.
Results can be evaluated by visual inspection (qualitative) or by densitometric data extraction (semi-quantitative).
Features
- One-step array processing (single incubation/single wash)
- Uses only 1-2 μl sample
- Entire experiment can be done within 1 hour
- Compatible with mouse sera, plasma, cell culture supernatants, or ascites fluids
- Sandwich ELISA-based detection imparts high specificity and sensitivity
- Low system CV, high reproducibility
- High throughput sample processing
- Qualitative visual inspection or semi-quantitative result
- Processed slides can be stored for years without signal decay
Mouse Ig Isotype Array Testing Service is available. For more details, click here.
Potential Applications
- Antibody-producing hybridoma screening and selection
- Selection and isolation of immunoglobulin isotype switch variants
- Mouse monoclonal antibody heavy and light chain identification
- Studying murine models of immunological diseases Monitoring biomarker levels in clinical trials
- Identifying potential molecular targets for drug development
Contents of Kits
- Mouse Isotyping Glass Chip
- Sample Diluent
- 20X Wash Buffer
- Labeled Detection Antibody
- Slide Washer/Dryer
- 96-well Plate
- Manual
Technical Notes
Data Analysis
The RayBio® Rapid Mouse Ig Isotyping Analyzer software (AAM-ISO-G1-SW) is available for one-step data com putation. An Excel-based program designed specifically for this array, the software tool outputs normalized digital values as well as isotype names. Simply copy and paste the densitometry values into the appropriate cells and the tool does the rest.
Specificity & Accuracy
The mouse isotype-specific capture antibodies in the kit have been tested to react only with their own isotyp; they exhibit no reactivity with other mouse Ig isotypes up to 1 μg/ml. The accuracy of the array is confirmed by isotype-predetermined mouse monoclonal antibody controls and third-party ELISA-determined hybridoma supernatant samples.
Sensitivity
The limit of detection (LOD) for each mouse Ig heavy or light chain has been determined with purified mouse immunoglobulin antigens to be in the nanogram range (see table below).
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