免疫磁珠说明书.doc

Contents 1. Description 1.1 Principle of the MACS? Separation 1.2 Background information 1.3 Applications 1.4 Reagent and instrument requirements试剂和仪器的要求 2. Protocol 2.1 Sample preparation样品制备 2.2 Magnetic labeling磁性标记 2.3 Magnetic separation磁性分离 3. Example of a separation using the CD133 MicroBead Kit 4. References 1. Description Components 2 mL CD133 MicroBeads, human:MicroBeads conjugated to monoclonal antihuman CD133 antibodies (isotype: mouse IgG1,clone AC133).2 mL FcR Blocking Reagent, human Specificity CD133 antigen, epitope (CD133/1)1.Capacity For 2亊10? total cells, up to 100 separations.Product format CD133 MicroBeads are supplied in buffercontaining stabilizer and 0.05% sodium azide.Storage Store protected from light at 2?8 °C. Do not freeze. The expiration date is indicated on the vial label. 1.1 Principle of the MACS? Separation First, the CD133+ cells are magnetically labeled with CD133 MicroBeads. Then, the cell suspension is loaded onto a MACS? Column, which is placed in the magnetic field of a MACS Separator. The magnetically labeled CD133+ cells are retained within the column. The unlabeled cells run through; this cell fraction is thus depleted of CD133+ cells. After removing the column from the magnetic field, the magnetically retained CD133+ cells can be eluted as the positively selected cell fraction. To increase the purity, the positively selected cell fraction containing the CD133+ cells is separated over a second column. 1.2 Background information The CD133 MicroBead Kit is a magnetic labeling system designed for the positive selection of CD133+ cells. It allows the single-step isolation of nonhematopoietic and early hematopoietic progenitors and stem cells. The CD133 molecule is a 5-transmembrane cell surface antigen with a molecular weight of 117 kD.2 The CD133/1 (clone AC133) antibody recognizes epitope 1 of the CD133 antigen.1 In the hematopoietic system, CD133 expression is restricted to a subset of CD34bright stem and progenitor cells in