To test LFA-1 expression, erythroleukemia cells (0.5 × 106 cells/run) were incubated with anti-human CD11a Alexa Fluor 594 clone HI111 (Biolegend) or anti-human CD11a FITC clone TS1/18 (Biolegend) (1:1000) for 1h at 25 °C and washed with PBS. FITC mouse IgG1 and Alexa Fluor 647 mouse IgG1 (Biolegend) served as isotype controls. To analyze cytosolic pH, K562 cells were labeled with pHrodo Green and treated with LtxA as for confocal imaging. At different periods of time (0, 15, and 30 min) the cells were analyzed by flow cytometry (509/533 nm). The fluorescence measurements were made with a BD LSR II flow cytometer (BD Biosciences). Ten thousand events were recorded per sample, and mean fluorescence intensities (MFI) were determined using WinList software.
Alexa fluor 647 mouse igg1
Manufactured by BioLegend
Sourced in United States
Alexa Fluor 647 mouse IgG1 is a fluorescently labeled mouse immunoglobulin G1 (IgG1) antibody. The antibody is conjugated to Alexa Fluor 647, a fluorescent dye with excitation and emission maxima at 650 nm and 665 nm, respectively. This product can be used for flow cytometry, immunofluorescence, and other immunoassay applications.
Automatically generated - may contain errors
Lab products found in correlation
Lsr 2 flow cytometer, by BD (1 mentions)
Fitc mouse igg1, by BioLegend (1 mentions)
Facscalibur, by BD (1 mentions)
Cell quest software 3, by BD (1 mentions)
Accuri c6, by BD (1 mentions)
Cd31 pe, by BD (1 mentions)
Cd45 fitc, by BioLegend (1 mentions)
Mouse igg2a fitc, by BioLegend (1 mentions)
Cd44 fitc, by BioLegend (1 mentions)
Cd90 apc, by BioLegend (1 mentions)
Pe mouse igg1, by BD (1 mentions)
3 protocols using alexa fluor 647 mouse igg1
1
LFA-1 Expression and Cytosolic pH Analysis
2
Flow Cytometric Analysis of EGFR Expression
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Stimulated or unstimulated A549 and PC-9 cells were stained with anti-human Alexa Fluor 647-coupled EGFR antibody or Alexa Fluor 647 mouse IgG1 (both Biolegend, San Diego, CA, USA) as previously described [33 (link)]. PBMC from co-cultures were stained with anti-human FITC-coupled CD3 antibody or FITC-coupled mouse IgG1 (both BD Biosciences) for 20 min. Subsequently, propidium iodide (10 μg/mL) was added for an additional 10 min. All samples were measured with flow cytometry (FACScalibur, BD Sciences, Heidelberg, Germany) and data were analyzed with Cellquest Software 3.0 (BD Sciences). Illustrations of the gating strategy can be found in Supplemental Figure S1 .
3
Characterization of Adipose-Derived Stem Cells
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MSC were validated by flow cytometry. Briefly, ADSC were detached and then washed once with FACS buffer (PBS + 0.5% BSA + 0.1% sodium azide). The cells were incubated with surface antibodies on ice for 20 min. After two times wash, cells were acquired by flow cytometry (Accuri C6, BD). The fluorescence conjugated antibodies for FACS include CD31-PE (Cat#555027, BD), CD44-FITC (203906, Biolegend), CD45-FITC (202205, Biolegend), and CD90-APC (202507, Biolegend). Matched isotype controls are as follows: FITC Mouse IgG2a, κ (400207, Biolegend); Alexa Fluor® 647 Mouse IgG1, κ (400130, Biolegend); and PE Mouse IgG1, κ (550617, BD).
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