b'Gel & Capillary | RUO AssaysGel and CapillaryIGH + IGK B-Cell Clonality AssaysResearch Use Only (RUO) AssayIGH + IGK B-Cell Clonality AssaysAssay UseIGH + IGK B-Cell Clonality Assays are useful for studies involving: The human immunoglobulin kappa (IGK) light chain locus on the short arm of chromosome 2 (2p11.2) spans 1820 kb. It is made up of Identification of clonal B-cell populations highly suggestive of76 variable (V) gene segments belonging to seven subgroups, five B-cell malignancies joining (J) gene segments, and one constant (C) gene segment. Lineage determination of leukemias and lymphomas Productive assembly of the kappa gene is successful in about 60% of Monitoring and evaluation of disease recurrence human B lymphocytes 2 ; however, even when unsuccessful, clonal B cells generally retain the rearranged kappa genes. The V segments Detection and assessment of residual disease encode the first 95 N-terminal amino acids. Positions 96-108 are Evaluation of new research and methods in malignancy studies encoded by one of five joining (J) gene segments. The constant (C) portion of the kappa light chain (amino acids 109-214) is encoded by a single constant (C) region separated from the J region by an intron.Summary and Explanation of the Test The length of the hypervariable CDR3 in kappa light chain genes is Five PCR master mixes are included in these test kits to test forlimited and rearrangements in this region display significant skewing rearrangements of both IGH and IGK. IGH Tubes A, B, and C target(platykurtosis). 3Therefore, clonal CDR3 products generated from this the conserved framework 1, 2, and 3 regions (respectively) within theregion are easily and reliably identified by heteroduplex analysis or variable (V H ) region and the joining (J H ) region of the IGH locus. IGKcapillary electrophoresis.tubes A and B target the variable (V), intragenic and joining (J), and kappa deleting element (K de ) regions of the IGK locus. Specimen RequirementsPositive and negative controls, as well as Specimen Control Size5 mL of peripheral blood, bone marrow biopsy, or bone marrow Ladder Master Mix are included. PCR products can be analyzed by capillary electrophoresis or heteroduplex analysis. Clonality isaspirate anti-coagulated with heparin or EDTA; or,indicated if any one of the master mixes generates clonal products. Minimum 5 mm cube of tissue; or,3 g of genomic DNA; or,BackgroundFormalin-fixed, paraffin-embedded (FFPE) tissue or slides.The immunoglobulin heavy chain (IGH) gene locus on chromosome 14 (14q32.33, formerly 14q32.3) includes 46-52 functional and 30Referencenonfunctional variable (V H ), 27 functional diversity (D H ), and 61.M Hummel et al., Leukemia 17: 2266-2272 (2003).functional joining (J H ) gene segments spread over 1250 kilobases. 1,2The most frequently used V Hgene segments in normal and malignant2. AW Langerak et al., Leukemia 17: 2272-2275 (2003).B cells belong to V H 3, V H 4, and V H 1 families, which together cover3. EP Rock, PR Sibbald, MM Davis, and YH Chien. J. Exp. Med. 179(1):7595% of V Husage. The V Hgene segments contain three framework323-328 (1994).regions (FR) and two complementarity determining regions (CDR).4. JJM van Dongen et al., Leukemia 17: 2257-2317 (2003).The FRs are characterized by their similarity among the various V Hsegments, whereas the CDRs are highly different even within the same V Hfamily. The CDRs represent the preferred target sequences for somatic hypermutations; however, somatic mutations can also occur in the FRs. Therefore, family-specific primers in the three different FRs were designed to increase the detection rate of clonal IGH B-cell populations and decrease the occurrence of false-negative results due to somatic hypermutation in primer binding sites. 1This assay is based on the EuroClonality/BIOMED-2 Concerted Action BMH4-CT98-3936.Figure Legend: Simple representation of the organization of a rearranged immunoglobulin heavy chain (IGH) gene on chromosome 14 and the immunoglobulin kappa light chain gene on chromosome 2p11.2. Black arrows represent the relativepositions of primers that target the conserved framework regions (FR1-3) and the downstream consensus J Hgene segments for IGH and the V, J, INTR and K deprimers which are included in the IGK master mix tubes.96 '