b'Gel & Capillary | RUO AssaysGel and CapillaryIGK Gene Clonality AssaysResearch Use Only (RUO) AssayIGK Gene Clonality AssaysAssay Useencode the first 95 N-terminal amino acids. Positions 96-108 are Immunoglobulin kappa light chain (IGK) gene clonality assays areencoded by one of five joining (J) gene segments. The constant (C) useful for the identification of B-cell clonality, studying clonal B-cellportion of the kappa light chain (amino acids 109-214) is encodedpopulations, and evaluation of new research and methods by a single constant (C) region separated from the J region in malignancy studies. by an intron. The length of the hypervariable complementarity determining region 3 (CDR3) in kappa light chain genes is limited and rearrangements in this region display significant skewing Summary and Explanation of the Test2(platykurtosis).Two master mixes target conserved regions within the variable(V1-7) and the joining (J1-5) regions that flank the uniqueTherefore, clonal CDR3 products generated from this region are hypervariable, antigen-binding, complementarity determiningmost easily and reliably identified by heteroduplex analysis using region 3 (CDR3). Other primers target the K deand intragenic regions.standard polyacrylamide gels. Alternatively, capillary electrophoresis or gene sequencing instruments coupled with differential fluorescence Tube A contains six upstream primers and two J region primers.detection can be used for analysis.Tube B contains six upstream V region primers, an upstream intragenic primer and a downstream K deprimer. Positive and negative controls, as well as a Specimen Control Size Ladder MasterSpecimen RequirementsMix, are included. PCR products can be analyzed by capillary electrophoresis or heteroduplex analysis. Clonality is indicated if anyThis assay tests extracted and purified genomic DNA (gDNA).one of the master mixes generates clonal products.BackgroundReferencesThe human immunoglobulin kappa (IGK) light chain locus on the1.AW Langerak et al., Leukemia 17: 2275-2280 (2003).2. EP Rock, PR Sibbald, MM Davis, and YH Chien. J. Exp. Med. 179(1): short arm of chromosome 2 (2p12, formerly 2p11.2) spans 1820 kb. It is made up of 76 variable (V) gene segments belonging to 7 subgroups,323-328 (1994).5 joining (J) gene segments, and one constant (C) gene segment.3. JJM van Dongen et al., Leukemia 17: 2257-2317 (2003).Productive assembly of the kappa gene is successful in about 60% of human B lymphocytes. 1However, even when unsuccessful, clonal B cells generally retain the rearranged kappa genes. The V segments This assay is based on the EuroClonality/BIOMED-2 Concerted Action BMH4-CT98-3936.Figure Legend: Schematic diagram of the immunoglobulin kappa light chain gene complex on chromosome 2p11.2. Shown are the relative positions and orientations for the V-J, and K deprimers, which are included in the IGK master mix tubes.104'