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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10⁹ 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