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).
Get tips on using CD326 (EpCAM) Monoclonal Antibody (G8.8), eFluor 450, eBioscience™ to perform Flow cytometry Anti-bodies Mouse - CD326/EpCAM
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Get tips on using CD137 (4-1BB) Monoclonal Antibody (17B5), eFluor 450, eBioscience™ to perform Flow cytometry Anti-bodies Mouse - CD137
Get tips on using CD80 (B7-1) Monoclonal Antibody (16-10A1), APC, eBioscience™ to perform Flow cytometry Anti-bodies Mouse - CD80
Get tips on using CD80 (B7-1) Monoclonal Antibody (16-10A1), PE, eBioscience™ to perform Flow cytometry Anti-bodies Mouse - CD80
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Get tips on using CD273 (B7-DC) Monoclonal Antibody (TY25), PE, eBioscience™ to perform Flow cytometry Anti-bodies Mouse - CD273/PD-L2
Get tips on using CD31 (PECAM-1) Monoclonal Antibody (390), Biotin, eBioscience™ to perform Flow cytometry Anti-bodies Mouse - CD31/Pecam-1
Get tips on using PCNA Antibody to perform Western blotting PCNA
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