Flow cytometry of MVs and cells was performed according to our published methods (Liu et al., 2007 (link); Li et al., 2010 (link), 2013 (link)). In brief, culture supernatants were fixed in filtered 1% paraformaldehyde (PFA), and supplemented with counting beads. The number of MVs in each sample was quantified using a FACSCalibur flow cytometer, with gating criteria based on particle size and surface exposure of PS, detected by PE-labeled annexin-V (BD Pharmingen) staining (Liu et al., 2007 (link); Li et al., 2010 (link), 2013 (link)). The portion of PS-positive MVs that were also HMGB1-positive was quantified by simultaneous staining with mouse anti-human HMGB1 antibody (R&D), followed by FITC-labeled goat anti-mouse IgG secondary antibody (Abcam). After treatment without or with TSE, adherent monolayers of macrophages were detached and fixed by suspension in 1% PFA for detection of cell-surface HMGB1 with flow cytometry (Li et al., 2013 (link)).
Fitc labeled goat anti mouse igg secondary antibody
Manufactured by Abcam
Sourced in China
FITC-labeled goat anti-mouse IgG secondary antibody is a laboratory reagent used to detect the presence of mouse immunoglobulin G (IgG) in samples. The antibody is conjugated with the fluorescent dye FITC (fluorescein isothiocyanate), which allows for the visualization of mouse IgG through fluorescence detection techniques.
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Lab products found in correlation
2 protocols using fitc labeled goat anti mouse igg secondary antibody
1
Flow Cytometry Analysis of Microvesicles
2
Fluorescent Labeling of T. reesei Transformants
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The T. reesei SUS2 and its DsRed-AfMP1 transformant spores were cultured in MM-lactose/sophorose (2% for lactose and 0.003% for sophorose, w/v) at 28 °C with agitation at 200 rpm for 12 h. The germinated spores were harvested by centrifugation at 10,000 rpm and washed twice with phosphate-buffered saline (PBS) containing 1 mg/ml bovine serum albumin (BSA). Cells were re-suspended in PBS containing 5 μg/ml of mouse anti-DsRed monoclonal antibody (Abcam, Shanghai, China) and incubated at room temperature for 2 h. The cells were washed again with PBS and incubated with 10 μg/ml of fluorescein isothiocyanate (FITC)-labeled goat anti-mouse IgG secondary antibody (Abcam) for 1 h. The cells were finally washed twice with PBS and passed through flow cytometry for analyses of both DsRed and FITC signals.
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