Imaging flow cytometry was performed using the ImageStreamX Mark II imaging flow cytometer (Amnis Corporation) equipped with a 40 × objective, 6 imaging channels, and 405 nm, 488 nm, and 642 lasers. For analysis of cell viability and CD45 expression, the enriched leukocytes were resuspended in 0.1% BSA in HEPES-buffered saline after RBC lysis and stained with the following antibodies and stains where applicable: DRAQ5 (1 μM; Cell Signaling Technologies), Sytox Blue (1 μM; Life Technologies), CellEvent Caspase-3/7 Green Detection Reagent (0.75 μM; Life Technologies), FITC-conjugated CD45 antibody (1:500; clone 5B1; Miltenyi Biotec), PE-conjugated CD66b antibody (1:125; clone G10F5; Stemcell Technologies), and PE-Cy7-conjugated CD16 antibody (1:200 or 1:333; clone 3G8; BD Biosciences). Single cells were gated using the nuclear marker DRAQ5. Neutrophils were identified by the dual positivity of CD66b and CD16. For analysis of neutrophil activation post-enrichment, cells were stained with DRAQ5 (1 μM; Cell Signaling Technologies), VioBlue-conjugated CD45 antibody (1:100; clone 5B1; Miltenyi Biotec), Alexa Fluor 488-conjugated CD11b antibody (1:500; clone ICRF44; Stemcell Technologies), PE-conjugated CD66b antibody (1:125; clone G10F5; Stemcell Technologies), and PE-Cy7-conjugated CD16 antibody (1:333; clone 3G8; BD Biosciences).
Fitc conjugated cd45 antibody
Manufactured by Miltenyi Biotec
Sourced in Germany
The FITC-conjugated CD45 antibody is a laboratory reagent used for the detection and identification of CD45-positive cells in various biological samples. CD45 is a transmembrane protein tyrosine phosphatase expressed on the surface of all nucleated hematopoietic cells. The FITC fluorescent label allows for the visualization and analysis of CD45-expressing cells using flow cytometry or other fluorescence-based detection methods.
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Lab products found in correlation
Draq5, by Cell Signaling Technology (1 mentions)
Pe conjugated cd66b antibody, by STEMCELL (1 mentions)
Alexa fluor 488 conjugated cd11b antibody, by STEMCELL (1 mentions)
Clone g10f5, by STEMCELL (1 mentions)
Pe cy7 conjugated cd16 antibody, by BD (1 mentions)
Cellevent caspase 3 7 green detection reagent, by Thermo Fisher Scientific (1 mentions)
Sytox blue, by Thermo Fisher Scientific (1 mentions)
2 protocols using fitc conjugated cd45 antibody
1
Multiparameter Imaging Flow Cytometry
2
Characterizing BM-MSC Phenotype Retention
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PE-conjugated CD44 and CD49 antibodies and FITC-conjugated CD45 antibody (from Miltenyi Biotec, Germany and Dako Co., Denmark, respectively) were used to perform flow cytometry analysis on the BM-MSCs. This was done to verify that the phenotype of the stem cells was retained after expanding in the cell culture.11 (link)
As the BM-MSCs were incubated, each of the antibodies was placed against the surface markers: for 4 min for the CD44 and CD90 antibodies and for 30 min for the CD45 antibody, all at 4°C. Afterwards, flow cytometry analysis was performed on the samples (Beckman Coulter Elite XL, Nyon, Switzerland).
The results of the flow cytometry analysis are tabulated inFigure 1 . Briefly, the analysis detected 76.88% of CD44, 85% of CD90, and a negligible 1.1% of CD45 in the isolated stem cells.
As the BM-MSCs were incubated, each of the antibodies was placed against the surface markers: for 4 min for the CD44 and CD90 antibodies and for 30 min for the CD45 antibody, all at 4°C. Afterwards, flow cytometry analysis was performed on the samples (Beckman Coulter Elite XL, Nyon, Switzerland).
The results of the flow cytometry analysis are tabulated in
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