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CAT #: 11020021

IGK Gene Clonality Assay for ABI Fluorescence Detection

Assay Use

This Research Use Only assay identifies immunoglobulin heavy chain clonality and is useful for the study of identifying clonal B-cell populations and evaluation of new research and methods in malignancy studies.

Product Details

  • Summary and Explanation of the Test

    BACKGROUND
    Rearrangements of the antigen receptor genes occur during ontogeny in B and T lymphocytes.  These gene rearrangements generate products that are unique in length and sequence for each cell.  Therefore, polymerase chain reaction (PCR) assays can be used to identify lymphocyte populations derived from a single cell by detecting the unique V-J gene rearrangements present within these antigen receptor loci.1  This PCR assay employs multiple consensus DNA primers that target conserved genetic regions within the immunoglobulin heavy chain gene.  This test is used to detect the vast majority of clonal B-cell malignancies from DNA.  Test products can be analyzed using a variety of detection formats, including gel and capillary electrophoresis.

    SUMMARY
    Invivoscribe’s gene clonality assays represent a simple approach to PCR-based clonality testing.  These standardized assays were carefully optimized testing positive and negative control samples using multiplex master mixes.

    This test kit includes three (3) master mixes.  The IGK Tube A master mix targets the variable (V) and the joining (J) regions of the Ig kappa light chain locus. Whereas the IGK Tube B master mix targets kappa deleting element (Kde) rearrangements with the variable (V) region and the intragenic Jϰ-Cϰ region.  The resulting Vϰ-Kde and Jϰ -Cϰ intron-Kde rearrangements are a result of unsuccessful rearrangements retained by the B-cell.  The third master mix, the Specimen Control Size Ladder, targets multiple genes and generates a series of amplicons of approximately 100, 200, 300, 400, and 600 base pairs (bp) to ensure that the quality and quantity of input DNA is adequate to yield a valid result.  A single thermal cycler program and similar detection methodologies are used with all of our Gene Clonality Assays.  This improves consistency and facilitates cross training on a broad range of different assays.

  • Specimen Requirements

    This assay tests genomic DNA.

References

1.  Rock, EP, et al. (1994) Journal of Experimental Medicine. Jan 1;179(1):323-8.

Disclaimer

This assay is based on the EuroClonality/BIOMED-2 Concerted Action BMH4-CT98-3936.

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