Gating strategies can be found in Supplemental Information (Figures S7 and S8 ). For analysis of mixed chimerism, cells were first stained with LIVE/DEAD Fixable Near-IR Dead Cell Stain Kit (ThermoFisher Scientific) and blocked with TruStain FcX anti-mouse (Biolegend) for 10 min on ice in Cell Stain Buffer (Biolegend). Antibodies used for staining from Biolegend were as follows: CD45.1 PerCp-Cy5.5 (A20), CD45.2 Pacific Blue (104), CD3 AF488 (17A2), CD4 PE (RM4–4), CD11b BV605 (M1/70), CD19 PE-Cy7 (6D5), CD49b APC (DX5), CD25 PE-Cy5 (PC61), CD8a PE (53–6.7), Ter-119 PE, CD11b PE (M1/70), Gr-1 PE (RB6–8C5), CD3 PE (17A2), B220 PE (RA3–6B2), CD317 PE (927), CD172a APC (P84), CD11c AF700 (N418), CD304 PE (3E12), FOXP3 AF647 (150D), Helios AF488 (22F6), B220 FITC (RA3–6B2), CD8a BV510 (53–6.7). eBioscience antibodies used were as follows: CD117 APC (2B8), Sca-1 Pe-Cy7 (D7). Staining of intracellular markers was conducted with Biolegend True-Nuclear Transcription Factor Buffer Set as per manufacturer’s instructions. For live cell sorting, propidium iodide (MilliporeSigma) was used to determine viability. Cells were analyzed and/or sorted with a BD FACSAria II. Data were analyzed using FlowJo (10.7).
Cd45 1 percp cy5
Manufactured by BioLegend
CD45.1 PerCp-Cy5.5 is a fluorochrome-conjugated antibody that binds to the CD45.1 allelic variant of the CD45 protein, which is a cell surface glycoprotein expressed on most hematopoietic cells. This product can be used for flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
Facsaria 2, by BD (2 mentions)
Sca 1 pe cy7, by Thermo Fisher Scientific (2 mentions)
Live dead fixable near ir dead cell stain kit, by Thermo Fisher Scientific (2 mentions)
Anti mouse trustain fcx, by BioLegend (2 mentions)
Cd3 af488, by BioLegend (2 mentions)
True nuclear transcription factor buffer set, by BioLegend (2 mentions)
Cell stain buffer, by BioLegend (2 mentions)
Cd45.2 pacific blue, by BioLegend (2 mentions)
Cd34 fitc, by Thermo Fisher Scientific (1 mentions)
Cd4 pe, by BioLegend (1 mentions)
B220 apc, by BioLegend (1 mentions)
C kit apc, by BioLegend (1 mentions)
Cd150 pe cy7, by BioLegend (1 mentions)
Cd8a pe, by BioLegend (1 mentions)
Cd11b apc, by BioLegend (1 mentions)
Cd135 pecy5, by BioLegend (1 mentions)
Ckit pe cy7, by BioLegend (1 mentions)
Cd48 apc cy7, by BioLegend (1 mentions)
Gr 1 pe, by BioLegend (1 mentions)
Sca 1 percp cy5, by Thermo Fisher Scientific (1 mentions)
3 protocols using cd45 1 percp cy5
1
Multiparameter Flow Cytometry Analysis of Hematopoietic Reconstitution
2
Multicolor Flow Cytometry Panel
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The following antibodies were used: mouse lineage cocktail-PE (BioLegend, cat# 78035), mouse lineage cocktail-APC (R&D Systems, cat# FLC001A), c-Kit-FITC (BioLegend, cat# 161603), c-Kit-APC (BioLegend, cat# 135108), c-Kit-PE/Cy7 (BioLegend, cat# 105814), c-Kit-BV421 (BioLegend, cat# 135124), Sca-1-BV605 (BioLegend, cat# 108133), Sca-1-APC (eBioscience, cat# 17-5981-82), Sca-1-BV421 (BioLegend, cat# 108127), Sca-1-PerCP-Cy5.5 (eBioscience, cat# 45-5981-82), CD150-BV421 (BioLegend, cat# 115925), CD150-PE (eBioscience, cat# 12-1501-82), CD150-PE-Cy7 (BioLegend, cat# 115913), CD48-PE/Cy7 (eBioscience, cat# 25-0481-80), CD48-BV711 (BioLegend, cat# 103439), CD48-APC-Cy7 (BioLegend, cat# 103432),CD34-FITC (eBioscience, cat# 11-0341-82), CD135-PE-Cy5 (BioLegend, cat# 135311), CD135-APC (BioLegend, cat# 135310), CD45.1-PerCP-Cy5.5 (BioLegend, cat# 110727), CD45.2-BV421 (BioLegend, cat# 109831), CD45.2-FITC(eBioscience, cat# 11-0454-82), Gr1-PE (BioLegend, cat# 108407), CD11b-APC (BioLegend cat# 101211), CD4-PE (BioLegend,cat# 116005), CD8a-PE (BioLegend, cat# 100707), B220-APC (BioLegend, cat# 103211).
3
Multiparameter Flow Cytometry Analysis of Hematopoietic Reconstitution
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Gating strategies can be found in Supplemental Information (Figures S7 and S8 ). For analysis of mixed chimerism, cells were first stained with LIVE/DEAD Fixable Near-IR Dead Cell Stain Kit (ThermoFisher Scientific) and blocked with TruStain FcX anti-mouse (Biolegend) for 10 min on ice in Cell Stain Buffer (Biolegend). Antibodies used for staining from Biolegend were as follows: CD45.1 PerCp-Cy5.5 (A20), CD45.2 Pacific Blue (104), CD3 AF488 (17A2), CD4 PE (RM4–4), CD11b BV605 (M1/70), CD19 PE-Cy7 (6D5), CD49b APC (DX5), CD25 PE-Cy5 (PC61), CD8a PE (53–6.7), Ter-119 PE, CD11b PE (M1/70), Gr-1 PE (RB6–8C5), CD3 PE (17A2), B220 PE (RA3–6B2), CD317 PE (927), CD172a APC (P84), CD11c AF700 (N418), CD304 PE (3E12), FOXP3 AF647 (150D), Helios AF488 (22F6), B220 FITC (RA3–6B2), CD8a BV510 (53–6.7). eBioscience antibodies used were as follows: CD117 APC (2B8), Sca-1 Pe-Cy7 (D7). Staining of intracellular markers was conducted with Biolegend True-Nuclear Transcription Factor Buffer Set as per manufacturer’s instructions. For live cell sorting, propidium iodide (MilliporeSigma) was used to determine viability. Cells were analyzed and/or sorted with a BD FACSAria II. Data were analyzed using FlowJo (10.7).
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