Cells were harvested and blocked with 2% bovine serum albumin (BSA; Sigma-Aldrich, B2064) for 20 min at room temperature. Then, the cells were stained with fluorescein-conjugated antibodies for 40 min at room temperature in 1% BSA. After incubation, cells were washed 3 times and analyzed with MoFlo (Beckman, Brea, CA, USA) and associated software. The antibodies used for flow cytometry were as follows: PE-conjugated mouse anti-human CD29 (Biolegend, San Diego, CA, USA; 303004), PE-conjugated mouse anti-human CD73 (BD Biosciences, San Jose, CA, USA; 550257), PE-conjugated mouse anti-human CD90 (eBioscience, San Diego, CA, USA; 12-0909-42), PE-conjugated mouse anti-human CD105 (Biolegend, 323206), PE-conjugated mouse anti-human CD34 (BD Biosciences, 555822), PE-conjugated mouse anti-human CD45 (BD Biosciences, 560957), PE-conjugated mouse anti-human HLA−DR (BD Biosciences, 555561), and the PE-conjugated mouse IgG1 (BD Biosciences, 551436) as an isotype control.
Pe conjugated mouse anti human cd45
Manufactured by BD
Sourced in United States
The PE-conjugated mouse anti-human CD45 is a laboratory reagent used for the detection and identification of human CD45-expressing cells in flow cytometry applications. It provides a specific and reliable means of identifying and quantifying cells expressing the CD45 antigen, which is a pan-leukocyte marker. This product is designed for research use only and its performance characteristics have been validated through standardized testing procedures.
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Lab products found in correlation
Pe conjugated mouse anti human cd73, by BD (2 mentions)
Pe conjugated mouse anti human cd34, by BD (1 mentions)
Pe conjugated mouse anti human cd90, by Thermo Fisher Scientific (1 mentions)
Pe conjugated mouse igg1, by BD (1 mentions)
Pe conjugated mouse anti human cd105, by BD (1 mentions)
Pe conjugated mouse anti human cd90, by BD (1 mentions)
Facscanto 2, by BD (1 mentions)
Anti human cd90 pe, by BioLegend (1 mentions)
Facs lsr 2, by BD (1 mentions)
Diva software, by BD (1 mentions)
3 protocols using pe conjugated mouse anti human cd45
1
Flow Cytometry Characterization of Mesenchymal Stem Cells
2
Flow Cytometric Analysis of hADSCs
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hADSCs were labeled with phycoerythrin (PE)-conjugated mouse anti-human CD73, PE-conjugated mouse anti-human CD90, PE-conjugated mouse anti-human CD105, and PE-conjugated mouse anti-human CD45 antibodies (BD Biosciences). Cells were then analyzed on a FACSCanto II (BD Biosciences) as described in the manufacturer’s protocol. A total of 10,000 events were acquired and the cells were gated properly for analysis.
3
Flow Cytometric Analysis of HBC Surface Markers
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HBC-specific surface markers were determined by flow cytometry. Four days after isolation, cells were detached by gently scraping them (Costar, New York, NY, USA) in HBSS.
Cell suspension was centrifuged with 300 g at 4°C for 5 min. Then, Fc receptors were blocked by incubation with 3% FBS/HBSS for 10 min at 4°C. PE-conjugated mouse anti-human CD45 (Cat#555483, BD Biosciences, Franklin Lakes, NJ, USA), V450-conjugated mouse anti-human CD80 (Cat#560442, BD), APC-conjugated mouse anti-human CD163 (Cat#333609, BioLegend), V450-conjugated mouse anti-human CD11b (Cat#560481, BD), PerCP-Cy5.5-conjugated mouse antihuman CD209 (#558263, BD), V450-conjugated mouse antihuman CD86 (Cat#560367, BD) and isotype-matched controls (Cat#345816, Cat#561504, Cat#345818, BD) were added and incubated for 30 min at 4°C. For identification of potential contaminating fibroblasts, PE anti-human CD90 (Cat#328110, BioLegend) was used. Afterward, cells were washed twice with PBS and resuspended in 150 µL PBS for analysis.
After setting the forward scatter threshold as to exclude cell debris, 10,000 events were collected using FACS LSRII and Diva software (BD). The results were analyzed using FlowJo software (Tree Star, Ashland, OR, USA). The percentage of positive cells was based on comparison with the isotypematched control antibody, for which gating was set at 1%.
Cell suspension was centrifuged with 300 g at 4°C for 5 min. Then, Fc receptors were blocked by incubation with 3% FBS/HBSS for 10 min at 4°C. PE-conjugated mouse anti-human CD45 (Cat#555483, BD Biosciences, Franklin Lakes, NJ, USA), V450-conjugated mouse anti-human CD80 (Cat#560442, BD), APC-conjugated mouse anti-human CD163 (Cat#333609, BioLegend), V450-conjugated mouse anti-human CD11b (Cat#560481, BD), PerCP-Cy5.5-conjugated mouse antihuman CD209 (#558263, BD), V450-conjugated mouse antihuman CD86 (Cat#560367, BD) and isotype-matched controls (Cat#345816, Cat#561504, Cat#345818, BD) were added and incubated for 30 min at 4°C. For identification of potential contaminating fibroblasts, PE anti-human CD90 (Cat#328110, BioLegend) was used. Afterward, cells were washed twice with PBS and resuspended in 150 µL PBS for analysis.
After setting the forward scatter threshold as to exclude cell debris, 10,000 events were collected using FACS LSRII and Diva software (BD). The results were analyzed using FlowJo software (Tree Star, Ashland, OR, USA). The percentage of positive cells was based on comparison with the isotypematched control antibody, for which gating was set at 1%.
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