C-kit-positive cells were isolated from the expanded cells using MACS, as described above. We performed a flow cytometry analysis to analyze the stem cell phenotypes. CSCs were isolated by enzymatic digestion using accutase, re-suspended in phosphate-buffered saline (PBS) and blocked with 3% fetal bovine serum for 15 min prior to the flow cytometry analysis. A nonspecific mouse IgG1 antibody (BD Biosciences, USA) was used as a control. Subsequently, CSCs were labeled with FITC-conjugated rat anti-mouse c-kit, FITC-conjugated rat anti-mouse spinocerebellar ataxia type 1 (Sca-1; BD Biosciences, USA), FITC-conjugated rat anti-mouse CD34 (Miltenyi Biotec Inc., Germany) and FITC-conjugated rat anti-mouse CD45 (Miltenyi Biotec Inc., Germany) antibodies for 30 min at 4 °C in the dark, washed twice with cold PBS, and then re-suspended in buffer. The data were obtained with a FACSCalibur flow cytometer (BD Biosciences, USA) and analyzed using WinMDI software.
Nonspecific mouse igg1 antibody
Manufactured by BD
Sourced in United States
Nonspecific mouse IgG1 antibody is a laboratory reagent used for general research purposes. It is a purified mouse immunoglobulin G1 (IgG1) antibody that does not have any specific target antigen. This antibody can be used as a control in various immunological experiments.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using nonspecific mouse igg1 antibody
1
Isolation and Characterization of Cardiac Stem Cells
2
Phenotypic Analysis of Stem Cells
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Flow cytometric analysis was performed to phenotypically analyze the stem cells were isolated from the expanded cells using MACS as described above. The CSCs were isolated by enzymatic digestion using Accutase(which did not affect surface markers of cells), washed in phosphate-buffered saline (PBS) and blocked with 3% fetal bovine serum for 15 min for flow cytometric analysis. A non-specific mouse IgG1 antibody (BD Biosciences, USA) was used as a control and the CSCs were labeled with FITC-conjugated rat anti-mouse c-kit (BD Biosciences, USA) for 30 min at 4°C in the dark. Subsequently, cells were washed twice with PBS solution and then re-suspended in buffer. Data were obtained with a FACS Calibur flow cytometer (BD Biosciences, USA) and analyzed by WinMDI software.
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