Megamix™, a mixture of monodisperse fluorescent beads of three diameters (0.5, 0.9 and 3 μm), was purchased from BioCytex (Marseille, France). Flow Cytometry Absolute Counting Standard microbeads (7.6 μm) were purchased from Bangs Laboratories, Inc. (Fishers, IN). Annexin V FITC, Glycophorin A-PE, Mouse IgG2bk-PE isotype control, and annexin V-binding buffer concentrate were purchased from BD Pharmingen (San Jose, CA).
Glycophorin a pe
Manufactured by BD
Sourced in United States
Glycophorin A-PE is a laboratory reagent used for the identification and enumeration of erythrocytes (red blood cells) in flow cytometric analyses. It is a fluorescently-labeled antibody that specifically binds to the Glycophorin A protein, which is a major sialoglycoprotein found on the surface of red blood cells. The PE (Phycoerythrin) label provides a fluorescent signal that can be detected by flow cytometry instruments.
Automatically generated - may contain errors
Lab products found in correlation
Cd45 apc, by BD (2 mentions)
Cd3 pe cy7, by BioLegend (2 mentions)
Cd15 bv605, by BD (2 mentions)
Cd14 bv605, by BioLegend (2 mentions)
Fceri pe, by BioLegend (2 mentions)
Cd19 percp cy5, by BioLegend (2 mentions)
Annexin 5 fitc, by BD (1 mentions)
Megamix, by Biocytex (1 mentions)
Cd33 bv421, by BioLegend (1 mentions)
Cd117 percpcy5, by BioLegend (1 mentions)
Cd33 pe cy7, by BD (1 mentions)
Lsr 2 flow cytometer, by BD (1 mentions)
Cd34 pe cy7, by BD (1 mentions)
Facsdiva software, by BD (1 mentions)
Cd45 pe, by BD (1 mentions)
Cd73 pe, by BD (1 mentions)
Hla dr fitc, by BD (1 mentions)
Cd31 apc, by BD (1 mentions)
Cd146 pe, by BD (1 mentions)
Facsaria, by BD (1 mentions)
4 protocols using glycophorin a pe
1
Fluorescent Bead-Based Assay Protocol
2
Multicolor Flow Cytometry Panel
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Cells were prepared for flow cytometry as previously described [3 (link)] and the following antibodies were used: CD45-APC, CD15-BV605, CD33-PECy7 and Glycophorin A-PE (BD Biosciences, San Jose, CA, USA), CD33-BV421, CD19-PerCPCy5.5, CD14-BV605, CD117-PerCPCy5.5, CD3-PECy7 and FceRI-PE (BioLegend, San Diego, CA, USA).
3
Multiparametric Phenotypic Profiling
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The following fluorochrome conjugated antibodies were used to label cell surface antigens: CD34 PE-Cy7, CD31 APC, CD73 PE, CD146 PE, CD45 PE, HLA DR FITC, and glycophorin A PE (BD Pharmingen). The relevant mouse isotypes (BD Pharmingen) were used as control. Cells were stained with labelled antibodies in the dark at 4°C for 45 min. A minimum of 30 000 events were acquired on a BD LSR II flow cytometer and the results were analyzed using BD FACSDiva software.
4
Multiparametric flow cytometry analysis
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Bones and spleens were crushed, cells resuspended and filtered to obtain a single-cell suspension that was analyzed by flow cytometry on a FACSCanto (BD Biosciences). Cells were stained with the following antibodies: CD45-APC, CD10-BV605, CD15-BV605 and glycophorin A-PE (from BD Biosciences) and CD33-BV421, CD19-PerCPCy5.5, CD34-APCCy7, CD68-PE, CD14-BV605, CD117-PECy7, IgM-PE, CD3-PECy7, FceRI-PE and CD25-BV421 (from BioLegend, San Diego, CA, USA). Control cells were stained with matching isotype controls. Sorting of cells was performed on a FACSAria (BD Biosciences).
For intracellular staining of phosphorylated STAT5 (signal transducer and activator of transcription 5), sorted pre-B cells were fixed in 1.6% paraformaldehyde for 10 min at room temperature. Cells were stored in 90% ethanol in −80 °C until analysis. Cells were washed two times in ice cold phosphate-buffered saline before resuspension in phosphate-buffered saline with 2% fetal calf serum. Cells were kept on ice and stained with antibodies against phosphorylated STAT5 (STAT5P-Alexa Flour 647 from BD) or matching isotype controls. Levels of phosphorylated STAT5 are presented as median fluorescence intensity, normalized to the median fluorescence intensity of isotype-stained cells.
For intracellular staining of phosphorylated STAT5 (signal transducer and activator of transcription 5), sorted pre-B cells were fixed in 1.6% paraformaldehyde for 10 min at room temperature. Cells were stored in 90% ethanol in −80 °C until analysis. Cells were washed two times in ice cold phosphate-buffered saline before resuspension in phosphate-buffered saline with 2% fetal calf serum. Cells were kept on ice and stained with antibodies against phosphorylated STAT5 (STAT5P-Alexa Flour 647 from BD) or matching isotype controls. Levels of phosphorylated STAT5 are presented as median fluorescence intensity, normalized to the median fluorescence intensity of isotype-stained cells.
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