Flow cytometry Anti-bodies Human - CD111/Nectin-1

Flow cytometry is an immunophenotyping technique whereby sing cell suspensions are stained for either cell surface markers or intracellular proteins by fluorescently-labelled antibodies and analyzed with a flow cytometer, where fluorescently-labelled molecules are excited by the laser to emit light at varying wavelengths, which is then detected by the instrument. There are several key criteria which are required to be kept in mind while designing a flow experiment- 1. Antibody titration (optimal dilution of antibodies should be calculated in order to avoid over- or under- saturated signals for proper detection of surface and intracellular markers), 2. Precision (3 or more replicates of the sample should be used per experiment), 3. Specificity (proper isotype controls should be included in the experiment), 4. Day-to-day variability (experiments should be repeated 3 or more times to ensure consistency and avoid variability due to flow cytometer settings), 5. Antibody interaction (Fluorescence minus one or FMO should be used, which is the comparison of signals from panel minus one antibody vs. the full panel), and 6. Antibody stability (fluorescently-labelled antibodies should be stored at 4C).

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Found 2 matching solutions for this experiment

Protocol tips
neurospheres were dissociated into a single cell suspension using Accutase (Innovative Cell Technologies) and prepared for fluorescence activated cell sorting (FACS) analysis
Downstream tips
Cells were analyzed using an LSR II Flow Cytometer or FACSCalibur (Becton Dickinson Biosciences) by the UAB Flow Cytometry Core Facility, and the results were expressed as a percentage of gated cells based on antibody binding using FlowJo version 10.0.6 software (Tree Star).
Protocol tips
1:100 for anti–nectin-1 antibody (Zymed)
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