b'Gel & Capillary | RUO AssaysGel and CapillaryIGH + IGK B-Cell Clonality AssaysResearch Use Only (RUO) AssayIGH + IGK B-Cell Clonality AssaysAssay Use 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 Immunoglobulin heavy chain (IGH) and Kappa light chain (IGK) gene76 variable (V) gene segments belonging to seven subgroups, five clonality assays are useful for identification of B-cell clonality, studyingjoining (J) gene segments, and one constant (C) gene segment. clonal B-cell populations, and evaluation of new research andProductive assembly of the kappa gene is successful in about 60% of methods in malignancy studies. human B lymphocytes 2 ; however, even when unsuccessful, clonal B cells generally retain the rearranged kappa genes. The V segments Summary and Explanation of the Testencode the first 95 N-terminal amino acids. Positions 96-108 are encoded by one of five joining (J) gene segments. The constant (C) Five PCR master mixes are included in these test kits to test forportion of the kappa light chain (amino acids 109-214) is encoded by a rearrangements of both IGH and IGK. IGH Tubes A, B, and C targetsingle constant (C) region separated from the J region by an intron.the conserved framework 1, 2, and 3 regions (respectively) within the variable (V H ) region and the joining (J H ) region of the IGH locus. IGK tubes A and B target the variable (V), intragenic and joining (J), andSpecimen Requirementskappa deleting element (K de ) regions of the IGK locus. This assay tests extracted and purified genomic DNA (gDNA)Positive and negative controls, as well as Specimen Control SizeReferencesLadder Master Mix are included. PCR products can be analyzed by capillary electrophoresis or heteroduplex analysis. Clonality is1.M Hummel et al., Leukemia 17: 2266-2272 (2003).indicated if any one of the master mixes generates clonal products. 2. AW Langerak et al., Leukemia 17: 2272-2275 (2003).3. EP Rock, PR Sibbald, MM Davis, and YH Chien. J. Exp. Med. 179(1):Background323-328 (1994).The immunoglobulin heavy chain (IGH) gene locus on chromosome4. JJM van Dongen et al., Leukemia 17: 2257-2317 (2003).14 (14q32.33, formerly 14q32.3) includes 46-52 functional and 30 nonfunctional variable (V H ), 27 functional diversity (D H ), and 6 functional joining (J H ) gene segments spread over 1250 kilobases. 1,2The most frequently used V Hgene segments in normal and malignant B cells belong to V H 3, V H 4, and V H 1 families, which together cover 7595% of V Husage. The V Hgene segments contain three framework regions (FR) and two complementarity determining regions (CDR). 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.98 '