Human Induced Pluripotent Stem Cell Analysis and Sorting

IPSC

The BD Stemflow™ Human iPSC Sorting and Analysis Kit contains a combination of mouse monoclonal antibody conjugates for the sorting and analysis of induced human pluripotent stem cell (hiPSC) cultures and cells in the process of being reprogrammed.  Following established reprogramming protocols, hiPSCs are sorted or analyzed from a heterogeneous cell culture utilizing the included antibody conjugates to three cell surface markers.  The antibody specificities that are provided in this kit and the cell populations they can identify are listed in the table below.  The kit also includes Isotype Controls and BD™ CompBead Plus.  Additional antibody formats that can enable more complex panels are available at www.bdbiosciences.com. Kit Components Component          Description                                                                                           Size                   Vol. Per Test                   Storage Buffer 51-9008175          PerCP-Cy™5.5 Mouse anti-Human CD13                                     50 Test                   5 µl                   Aqueous buffered solution containing                                                                                                                                                                                     BSA and ≤0.09% sodium azide 51-9008176          Alexa Fluor® 647 Mouse anti-SSEA-4                                              50 Test                   5 µl                   Aqueous buffered solution containing                                                                                                                                                                                     BSA and ≤0.09% sodium azide 51-9008177          PE Mouse anti-Human TRA-1-60 Antigen                                     50 Test                   5 µl                   Aqueous buffered solution containing                                                                                                                                                                                     protein stabilizer, BSA and ≤0.09%                                                                                                                                                                                                                                                                                                                                       sodium azide 51-9008178          PerCP-Cy™5.5 Mouse IgG1, κ Isotype Control                            50 Test                   5 µl                   Aqueous buffered solution containing                                                                                                                                                                                     BSA and ≤0.09% sodium azide 51-9008179          Alexa Fluor® 647 Mouse IgG3, κ Isotype Control                   50 Test                   5 µl                   Aqueous buffered solution containing                                                                                                                                                                                     BSA and ≤0.09% sodium azide 51-9008180          PE Mouse IgM, κ Isotype Control                                                       50 Test                   5 µl                   Aqueous buffered solution containing                                                                                                                                                                                     BSA and ≤0.09% sodium azide 51-9006227          Negative Control (PBS with 1% BSA) CompBead Plus          6 ml                            1 drop          Aqueous buffered solution containing                                                                                                                                                                                     BSA and ≤0.09% sodium azide 51-9006274          Anti-Mouse Ig, κ CompBead Plus                                                       6 ml                            1 drop          Aqueous buffered solution containing                                                                                                                                                                                     BSA and ≤0.09% sodium azide Specificity          Clone                            Cell population identified CD13                   WM15                            Fibroblasts, cells that are not reprogrammed TRA-1-60          TRA-1-60.1                   Pluripotent stem cells SSEA-4                   MC-813-70.1                   Pluripotent stem cells

The BD Stemflow™ Human iPSC Sorting and Analysis Kit contains a combination of mouse monoclonal antibody conjugates for the sorting and analysis of induced human pluripotent stem cell (hiPSC) cultures and cells in the process of being reprogrammed.  Following established reprogramming protocols, hiPSCs are sorted or analyzed from a heterogeneous cell culture utilizing the included antibody conjugates to three cell surface markers.  The antibody specificities that are provided in this kit and the cell populations they can identify are listed in the table below.  The kit also includes Isotype Controls and BD™ CompBead Plus.  Additional antibody formats that can enable more complex panels are available at www.bdbiosciences.com. Kit Components Component          Description                                                                                           Size                   Vol. Per Test                   Storage Buffer 51-9008175          PerCP-Cy™5.5 Mouse anti-Human CD13                                     50 Test                   5 µl                   Aqueous buffered solution containing                                                                                                                                                                                     BSA and ≤0.09% sodium azide 51-9008176          Alexa Fluor® 647 Mouse anti-SSEA-4                                              50 Test                   5 µl                   Aqueous buffered solution containing                                                                                                                                                                                     BSA and ≤0.09% sodium azide 51-9008177          PE Mouse anti-Human TRA-1-60 Antigen                                     50 Test                   5 µl                   Aqueous buffered solution containing                                                                                                                                                                                     protein stabilizer, BSA and ≤0.09%                                                                                                                                                                                                                                                                                                                                       sodium azide 51-9008178          PerCP-Cy™5.5 Mouse IgG1, κ Isotype Control                            50 Test                   5 µl                   Aqueous buffered solution containing                                                                                                                                                                                     BSA and ≤0.09% sodium azide 51-9008179          Alexa Fluor® 647 Mouse IgG3, κ Isotype Control                   50 Test                   5 µl                   Aqueous buffered solution containing                                                                                                                                                                                     BSA and ≤0.09% sodium azide 51-9008180          PE Mouse IgM, κ Isotype Control                                                       50 Test                   5 µl                   Aqueous buffered solution containing                                                                                                                                                                                     BSA and ≤0.09% sodium azide 51-9006227          Negative Control (PBS with 1% BSA) CompBead Plus          6 ml                            1 drop          Aqueous buffered solution containing                                                                                                                                                                                     BSA and ≤0.09% sodium azide 51-9006274          Anti-Mouse Ig, κ CompBead Plus                                                       6 ml                            1 drop          Aqueous buffered solution containing                                                                                                                                                                                     BSA and ≤0.09% sodium azide Specificity          Clone                            Cell population identified CD13                   WM15                            Fibroblasts, cells that are not reprogrammed TRA-1-60          TRA-1-60.1                   Pluripotent stem cells SSEA-4                   MC-813-70.1                   Pluripotent stem cells

Flow cytometry (Routinely Tested)

Human (QC Testing)

研发参考(5) 1. Chan EM, Ratanasirintrawoot S, Park IH, et al. Live cell imaging distinguishes bona fide human iPS cells from partially reprogrammed cells. Nat Biotechnol. 2009; 27(11):1033-1037. (Methodology: Cell culture, Fluorescence microscopy). 2. Park IH, Arora N, Huo H, et al. Disease-specific induced pluripotent stem cells. Cell. 2008; 134(5):877-886. (Biology). 3. Robinton DA, Daley GQ. The promise of induced pluripotent stem cells in research and therapy. Nature. 2012; 481(7381):295-305. (Biology). 4. Takahashi K, Yamanaka S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell. 2006; 126(4):663-676. (Biology). 5. Valamehr B, Abujarour R, Robinson M, et al. A novel platform to enable the high-throughput derivation and characterization of feeder-free human iPSCs. Sci Rep. 2(213)(Methodology: Cell separation, Flow cytometry).