b'Gel & Capillary | RUO AssaysGel and Capillary Gel and CapillaryIGH Gene Clonality AssaysResearch Use Only (RUO) Assay IGH Gene Clonality AssaysResearch Use Only (RUO) AssayIGH Gene Clonality AssaysAssay Use The FRs are characterized by their similarity among the various V H Reagentssegments, whereas the CDRs are highly different even within the sameControls Concentration Units in Assay Units in Assay MegaKitImmunoglobulin heavy chain (IGH) clonality assays are useful forV Hfamily. The CDRs represent the preferred target sequences for identification of B-cell clonality, studying clonal B-cell populations, andsomatic hypermutations; however, somatic mutations can also occurIVS-0030Clonal Control DNA 200 g/mL 1 x 100 L tube 5 x 100 L tubesevaluation of new research and methods in malignancy studies. in the FRs. Therefore, family-specific primers in the three differentIVS-0019 Clonal Control DNA 200 g/mL 1 x 100 L tube 5 x 100 L tubesFRs were designed to increase the detection rate of clonal IGH B-cell populations and decrease the occurrence of false-negative resultsIVS-0024 Clonal Control DNA 200 g/mL 1 x 100 L tube 5 x 100 L tubesSummary and Explanation of the Test1due to somatic hypermutation in primer binding sites. In addition toIVS-0008 Clonal Control DNA 200 g/mL 1 x 100 L tube 5 x 100 L tubesFive master mixes target conserved regions within the variable (V H ),V H -J Hrearrangements, incomplete D H -J Hrearrangements have been diversity (D H ), and the joining (J H ) regions that flank the uniquefound in mature and immature B-cell malignancies. Therefore, D H -J H IVS-0000 Polyclonal Control DNA 200 g/mL 1 x 100 L tube 5 x 100 L tubeshypervariable, antigen-binding, complementarity determiningPCR analysis may be of added value for clonality assessment. 2 Master Mixes Target Units in Assay Units in Assay MegaKitregion 3 (CDR3). Tube A contains six framework region 1 (FR1)IGH Tube A Framework 1 + J H 1 x 1500 L tube 10 x 1500 L tubesprimers and a consensus J Hregion primer. Tube B contains seven framework region 2 (FR2) primers and a consensus J Hprimer. TubeIGH Tube B Framework 2 + J H 1 x 1500 L tube 10 x 1500 L tubesC contains seven framework region 3 (FR1) primers and a consensusSpecimen RequirementsJ Hprimer. Tube D contains six D Hregion primers and a consensus J H IGH Tube C Framework 3 + J H 1 x 1500 L tube 10 x 1500 L tubesregion primer. Tube E contains a D H 7 region primer and a consensusThis assay tests extracted and purified genomic DNA (gDNA). IGH Tube D D H 1-6 + J H 1 x 1500 L tube 10 x 1500 L tubesJ Hprimer. Positive and negative controls, as well as the Specimen Control Size Ladder Master Mix are included. PCR products canIGH Tube E D H 7 + J H 1 x 1500 L tube 10 x 1500 L tubesbe analyzed by capillary electrophoresis or heteroduplex analysis.References Specimen Control Size Ladder Multiple Genes 1 x 1500 L tube 10 x 1500 L tubesClonality is indicated if any one of the master mixes generates a1.M Hummel et al., Leukemia 17:2266-2272 (2003).clonal product. 2. AW Langerak et al., Leukemia 17:2272-2275 (2003).3. JJM van Dongen et al., Leukemia 17:2257-2317 (2003).BackgroundOrdering InformationThe immunoglobulin heavy chain (IGH) gene locus on chromosomeCatalog # Products Quantity14 (14q32.33, formerly 14q32.3) includes 46-52 functional and 30 nonfunctional variable (V H ), 27 functional diversity (D H ), and 61-101-0020 IGH Gene Clonality Assay - Gel Detection 33 reactionsfunctional joining (J H ) gene segments spread over 1250 kilobases. 1,2The1-101-0040 IGH Gene Clonality Assay MegaKit - Gel Detection330 reactionsmost frequently used V Hgene segments in normal and malignant B cells belong to the V H 3, V H 4, and V H 1 family, together covering 7595%1-101-0061IGH Gene Clonality Assay - ABI Fluorescence Detection 33 reactionsof V Husage. The V Hgene segments contain three framework regions1-101-0081 IGH Gene Clonality Assay MegaKit - ABI Fluorescence Detection330 reactions(FR) and two complementarity determining regions (CDR).This 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 gene on chromosome 14. Black arrows represent the relative positions of primers that target the conserved framework (FR1-3) and diversity (D H 1-7) regions, and the downstream consensus J Hgene segments. The amplicon products generated from each of these regions can be differentially detected when fluorescent primer sets are used with capillary electrophoresis instruments that employ differential fluorescence detection.100Invivoscribe 2021|101'