50 μL peripheral blood or ~5×105 bone marrow cells in PBS with 10% FBS were blocked with anti-mouse CD16/CD32 (cat # 101302, Biolegend, San Diego, CA, USA) and then stained with fluorophore conjugated antibodies. Following staining, red blood cells were lysed with RBC lysis buffer (cat # sc-296258, Santa Cruz Biotechnology, Dallas, TX, USA) for 10 min and washed twice with 10% FBS in PBS. Peripheral blood samples underwent a second 5 min RBC lysis prior to washing. Samples were acquired on a BD Accuri C6 or Beckman coulter Gallios flow cytometer and analyzed using BD Accuri Csampler software or FlowJo. Cells were stained with anti-human CD34 PE (cat # 343606, Biolegend), anti-human CD11b PE (cat # 555388, BD Biosciences, San Jose, CA, USA), anti-human CD3 PE/Dazzle™ 594 (cat # 300450, Biolegend), anti-mouse CD45 PerCP-Cy5.5 (cat # 103132 Biolegend), anti-mouse CD45 PerCP (cat # 557235, BD), anti-human CD19 PE-Cy7 (cat # 302216, Biolegend), anti-human CD45 APC (cat # 555485, BD Bioscience or cat # 304012, Biolegend). Analysis based 10 000 bone marrow and 3 000 blood cells from the live cell gate.
Anti mouse cd45 percp
Manufactured by BD
Sourced in United States
The Anti-mouse CD45 PerCP is a fluorescent-labeled antibody that binds to the CD45 antigen expressed on the surface of mouse leukocytes. It is used in flow cytometry applications to identify and quantify different leukocyte populations.
Automatically generated - may contain errors
Lab products found in correlation
Anti human cd11b pe, by BD (2 mentions)
Anti human cd19 pe cy7, by BioLegend (2 mentions)
Anti mouse cd16 cd32, by BioLegend (2 mentions)
Rbc lysis buffer, by Santa Cruz Biotechnology (2 mentions)
Anti mouse cd45 percp cy5, by BioLegend (2 mentions)
Anti human cd45 apc, by BD (2 mentions)
Anti human cd34 pe, by BioLegend (2 mentions)
Anti human cd3 pe dazzle 594, by BioLegend (2 mentions)
Facsdiva software, by BD (1 mentions)
Cd3 fitc, by BD (1 mentions)
Cd19 fitc, by BD (1 mentions)
Cd19 apc h7, by BD (1 mentions)
Annexin 5 binding buffer, by Thermo Fisher Scientific (1 mentions)
Facs fortessa flow cytometer, by BD (1 mentions)
Cd15 fitc, by BD (1 mentions)
Flow count fluorosphere beads, by Beckman Coulter (1 mentions)
Anti mouse mhcii pe, by BD (1 mentions)
Anti mouse cd80 apc, by BD (1 mentions)
4 protocols using anti mouse cd45 percp
1
Multiparameter Flow Cytometry of Hematopoietic Cells
2
Multiparameter Flow Cytometry of AML Cells
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AML cell lines and primary cells were treated with CPI-637, NEO1132, or NEO2734 for 96 hours before analysis. Cells were harvested and washed in PBS with 0.1% human serum albumin (PBS/HSA). Samples were incubated for 15-30 minutes at room temperature with monoclonal antibody combinations consisting of antihuman 7AAD-PerCP (1:10), CD45-HV500c (2D1, 1:20), CD33-PE (1:20) or CD33-PC7 (1:20), CD34-APC (8G12, 1:50) or CD34-BV421 (581, 1:20), CD38-APC (HB7, 1:50), CD15-FITC (HI98, 1:100), CD7-APC (M-T701, 1:50) or CD7-FITC (M-T701, 1:20), CD56-PE (MY31, 1:20), HLA-DR-FITC (L243, 1:100), CD3-FITC (SK7, 1:50) or CD3-PE (1:50), CD19-APC-H7 (SJ25C1, 1:10) or CD19-FITC (89B-B4, 1:20), or antimouse CD45-PerCP (30-F11, 1:50), from BD Biosciences, and washed once with PBS/HSA. For detection of apoptosis, cells were stained with 7AAD-PerCP (1:10) for 15-30 minutes, washed with PBS/HSA, and subsequently stained with AnnexinV-FITC 1:1000 (Tau Technologies) in AnnexinV-binding buffer (Invitrogen) for 15 minutes on ice. Flow-count fluorosphere beads (Beckman Coulter, Brea) were added according to manufacturer instruction directly before analysis using a FACS-Fortessa flow cytometer (BD Biosciences). Data analysis was performed with FACS Diva software (BD Biosciences).
3
Multiparameter Flow Cytometry of Hematopoietic Cells
4
Medullary Phenotype Analysis of AIRE Spheroids
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We used a BD FACSymphony™ A1 Cell Analyzer (Becton Dickinson, Franklin Lakes, NJ) flow cytometer to evaluate the medullary phenotype of AIREWTor AIRE−/− spheroids. A sample of 1 × 106 mTECs separated from spheroids were labeled with 1:250 dilution of anti-mouse CD45-PerCP, anti-mouse CD326 (EpCam), anti-mouse Ly51-PE (BD Biosciences, San Jose, CA), anti-mouse CD80-APC, and anti-mouse MHCII-PE antibodies in a final volume of 200 μL of cell suspension. Labeling was also performed with a 1:250 dilution of Lectin agglutinin I (Ulex europaeus UEA-I)-FITC (Vector Labs, Burlingame, CA) and the viability marker DAPI- BD Pharmingen™ .
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