In brief, to block nonspecific binding, each cell line was resuspended in PBS after being washed twice with PBS, and incubated with human IgG antibody (#I2511, SIGMA) on ice for 15 min. After blocking, the cells (0.5 million cells/50 μL) were incubated on ice in the dark for 30 min with 5 μL/sample of phycoerythrin (PE)‐conjugated rat antihuman CD49f (#313612, BioLegend, 100 μg/mL), fluorescein isothiocyanate (FITC)‐conjugated mouse antihuman CD29 (#6603109, Beckman Coulter, 2 mg/mL), or PE‐conjugated mouse antihuman CD104 (#327808, BioLegend, 200 μg/mL). As isotype controls, the other aliquots of cells were incubated with PE‐conjugated rat IgG2b,κ (#553989, BD Pharmingen) or mouse IgG1 (#A07796, Beckman Coulter). After washing twice with ice‐cold PBS, the cells were analyzed using flow cytometry (FACSCalibur, BD Biosciences, San Jose, CA).
Mouse igg1
Manufactured by Beckman Coulter
Mouse IgG1 is a type of immunoglobulin G1 antibody derived from mouse. It is commonly used as a control or reference antibody in various laboratory applications.
Automatically generated - may contain errors
2 protocols using mouse igg1
1
Flow Cytometric Analysis of Integrin Expression
2
Flow Cytometric Characterization of Adipose-Derived Mesenchymal Stem Cells
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The cAd-MSCs phenotypes were evaluated by performing flow cytometry. The cells were pelleted at 1500 rpm for 5 min, washed with MACS buffer (Miltenyi Biotec), and then distributed into groups according to requirement. Here we had four markers including CD34(R-Phycoerythrin conjugated, clone 1H6; BD), CD44(ALEXA FLUOR® 488-conjugated, clone YKIX337.8.7; Bio-Rad), CD45(R-Phycoerythrin conjugated clone YKIX337.8.7; Bio-Rad), and CD90(clone YKIX337.8.7; Bio-Rad). Apart from CD90, other groups were immunostained for 30 min at 4°C in the dark. CD90 requires another 30 min of incubation for secondary antibody (R-Phycoerythrin conjugated, Rabbit anti-rat, Bio-Rad). Furthermore, there were three relative isotype control antibodies that need to be prepared, including Rat igG2a (ALEXA FLUOR® 488 isotype control, Bio-Rad), Rat igG2b (R-Phycoerythrin isotype control, Bio-Rad), and Mouse igG1 (R-Phycoerythrin isotype control, Beckman Coulter), which were also incubated 30 min at 4°C in the dark. After incubation, the cells were washed and resuspended with MACS buffer. The acquisition was performed by using a FACScan (BECTON DICKINSON) with 5,000 events per sample. Lastly, data were analyzed by using FlowJo software (Tree Star).
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