EVs samples were adsorbed onto 4% w/v, 4-µm aldehyde/sulfate latex beads (Thermo Fisher Scientific) for 1 h at 4 °C. The ratio between the number of beads and EVs was kept constant (1:1) for all samples. The reaction was stopped by adding 100 mM glycine and washed at 800g with 1 ml of PBS 1X. Membrane-bound beads were incubated with mouse anti-CD63 (1:100, 556019, BD-Bioscience) for 1 h at RT and stained with a FITC-conjugated secondary antibody (1:200, R&D Systems) for 1 h at RT. After a final washing step, the samples were resuspended in 0.5 ml of PBS 1X and analyzed using the Attune Acoustic Focusing Cytometer (Thermo Fisher Scientific).
Fitc conjugated secondary antibody
Manufactured by R&D Systems
Sourced in United Kingdom
FITC-conjugated secondary antibody is a laboratory reagent used to detect the presence of primary antibodies in various immunoassays. It consists of a fluorescent dye, fluorescein isothiocyanate (FITC), which is covalently attached to a secondary antibody molecule. This allows for the visualization and localization of target antigens or proteins that have been bound by the primary antibody.
Automatically generated - may contain errors
Lab products found in correlation
Attune acoustic focusing cytometer, by Thermo Fisher Scientific (1 mentions)
4 mm aldehyde sulphate latex beads, by Thermo Fisher Scientific (1 mentions)
Mouse anti cd63, by Abcam (1 mentions)
Facscalibur flow cytometer, by BD (1 mentions)
Hep3b, by American Type Culture Collection (1 mentions)
Facsaria cell sorter, by BD (1 mentions)
Pe conjugated anti cd11b, by BioLegend (1 mentions)
Af4670, by R&D Systems (1 mentions)
Hepg2, by R&D Systems (1 mentions)
3 protocols using fitc conjugated secondary antibody
1
Characterization of Extracellular Vesicles
2
Extracellular Vesicle Characterization by Flow Cytometry
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AB, SMV, or EXO samples were adsorbed onto 4-mm aldehyde/sulphate latex beads (Invitrogen, Paisley, UK) overnight at 4°C. The reaction was stopped by adding glycine 100 mM. Membrane-bound beads were washed in PBS/1% BSA, incubated with mouse anti-CD63 (Abcam, Cambridge, UK) or appropriate isotype control for 30 min at 4°C, stained with FITC-conjugated secondary antibody (R&D Systems) for 30 min at 4°C, and resuspended in 0.5 ml of PBS. Samples were analyzed using a FACS Calibur flow cytometer (BD Biosciences, San Jose, CA, USA).
3
CD47 Expression in Hepatocyte Cell Lines
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