BD BCR-ABL Protein Kit For determining BCR-ABL fusion proteins Based on the BD™ CBA system, this kit About fusion proteins detects the presence of specific fusion Fusion proteins are a consequence of chromosomal rearrangements in a cell. Some chromosomal rearrangements proteins in cell lysates. lead to changes in the positions of genes, which ultimately cause the cells to generate altered proteins. Fusion proteins are absent from normal, healthy cells. Fusion proteins have antigen determinants derived from each of Features the original proteins that would not normally be present on the No operator-dependent data interpretation same molecular structure. By introducing two antibodies into a sample, each specific to an antigen on a different portion, it is Flow cytometry based immunoassay possible to detect the presence of a fusion protein. Principle of the assay Specific to fusion proteins in cell lines, sensitive to K562 cell concentrations as low as 10% Flow cytometry allows for the discrimination of particles on the basis of attributes such as size and fluorescence. BD Cytometric Faster results than existing assays Bead Array (CBA) systems provide a way of coupling a soluble analyte or set of analytes with beads of known size and fluorescence, making it possible to detect analytes through flow cytometry.1 The BD™ BCR-ABL Protein Kit uses this technology to detect BCR-ABL fusion proteins2 in human blood research samples. Figure 1. Once cell lysates are prepared using the BD BCR-ABL Protein Kit and samples are run in the flow cytometer, histograms show the pres- ence or absence of BCR-ABL fusion proteins. bdbiosciences.com BD BCR-ABL Protein Kit About the assay kit References 1. Morgan E, Varro R, Sepulveda H, et al. Cytometric The BD BCR-ABL Protein Kit, a bead- These sandwich complexes can be bead array: a multiplexed assay platform with appli- based immunoassay, was developed studied by using flow cytometry to cations in various areas of biology. Clin Immunol. using the BD Cytometric Bead Array identify particles with attributes of both 2004;110:252-266. (CBA) system. BD CBA assays take the bead and the detector. 2. Ben-Neriah Y, Daley GQ, Mes-Masson AM, advantage of flow cytometry’s excellent Witte ON, Baltimore D. The chronic myelogenous Kit contents leukemia-specific P210 protein is the product of the fluorescence detection capabilities. Key bcr/abl hybrid gene. Science. 1986;233:212-214. components in the kit are the capture • Pretreatment A (powder), store at –20°C. beads, which are stained particles coated with an antibody, and the detector • Pretreatment B (powder), store at 2°C antibodies, which are conjugated to a to 8°C. fluorescent dye. • Capture beads, store in the dark at The capture beads included in this kit 2°C to 8°C. have been coated with an antibody • Detector reagent, store in the dark at specific to one part of the BCR-ABL 2°C to 8°C. fusion protein. The detector reagent is a phycoerythrin (PE)–conjugated antibody specific to a different part of the protein. The capture beads and detector reagent are incubated with a prepared sample. If the sample contains BCR-ABL proteins, sandwich complexes are formed, as shown in Figure 2. Figure 2. Formation of sandwich complexes in the BCR-ABL immunoassay. Ordering Information Description Size Cat.No. BD BCR-ABL Protein Kit 50 tests 643939 BD BCR-ABL Buffer Kit 100 tests 560272 BD Biosciences POB 13, Erembodegem-Dorp 86 B-9320 Erembodegem Belgium Tel.: (32) 2 400 98 95 For Research Use Only. Not for use in diagnostic or therapeutic procedures. BD, BD Logo and all other trademarks are property of Becton, Dickinson and Company. © 2008 BD Fax: (32) 2 401 70 94 XEUR0221-00 bdbiosciences.com.