Flow cytometry Anti-bodies Human - CD10

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 3 matching solutions for this experiment

Purified Mouse Anti-Human CD10

BD Biosciences

Upstream tips	Protocol tips	Downstream tips
	100 µL of the test sample (fresh bone marrow) was added in all tubes. The cells were incubated for 20 min at room temperature (18–25°C) in dark. The RBCs were lysed using 2 ml of 1× BD FACS lysing solution (BD Biosciences, San Jose, CA, USA). The above tubes were centrifuged for 5 min at 1800 g at room temperature. Supernatant was discarded and the pellet was re-suspended. The pellet was washed twice with 2 ml of sheath fluid (FACSFlow, Becton Biosciences, San Jose, CA, USA) at 1800 g for 5 min.	The pellet was obtained after washing and was resuspended in 300 µl of sheath fluid in the tube and a total of 20,000 events were acquired. The samples were then analyzed using a six-color flow cytometer (BD FACSVerse, Becton Biosciences, San Jose, CA, USA)

Protocol tips
100 µL of the test sample (fresh bone marrow) was added in all tubes. The cells were incubated for 20 min at room temperature (18–25°C) in dark. The RBCs were lysed using 2 ml of 1× BD FACS lysing solution (BD Biosciences, San Jose, CA, USA). The above tubes were centrifuged for 5 min at 1800 g at room temperature. Supernatant was discarded and the pellet was re-suspended. The pellet was washed twice with 2 ml of sheath fluid (FACSFlow, Becton Biosciences, San Jose, CA, USA) at 1800 g for 5 min.
Downstream tips
The pellet was obtained after washing and was resuspended in 300 µl of sheath fluid in the tube and a total of 20,000 events were acquired. The samples were then analyzed using a six-color flow cytometer (BD FACSVerse, Becton Biosciences, San Jose, CA, USA)

Manufacturer protocol Publication protocol 1 Relevant paper

PE-Cy™7 Mouse Anti-Human CD10

BD Biosciences

Upstream tips	Protocol tips	Downstream tips
	Briefly, approximately 100 μL of EDTA‐anticoagulated pleural fluid samples were immunophenotyped using six combinations of monoclonal antibodies and a direct immunofluorescence stain.	The cells were acquired in a FACSCanto flow cytometer (Becton Dickinson, San Jose, CA) using the FACSDiva software (Becton Dickinson). A minimum of 100,000 events were acquired. For data analysis the Infinicyt software (Cytognos SL, Salamanca, Spain) was used.

Protocol tips
Briefly, approximately 100 μL of EDTA‐anticoagulated pleural fluid samples were immunophenotyped using six combinations of monoclonal antibodies and a direct immunofluorescence stain.
Downstream tips
The cells were acquired in a FACSCanto flow cytometer (Becton Dickinson, San Jose, CA) using the FACSDiva software (Becton Dickinson). A minimum of 100,000 events were acquired. For data analysis the Infinicyt software (Cytognos SL, Salamanca, Spain) was used.

Manufacturer protocol Publication protocol 1 Relevant paper

PE Mouse Anti-Human CD10

BD Biosciences

Upstream tips	Protocol tips	Downstream tips
	Heparinized bone marrow (BM) and peripheral blood (PB) cells (106 leukocytes per tube) were incubated with various combinations of mAb (Supplementary Table S3) for 15 minutes.

Protocol tips
Heparinized bone marrow (BM) and peripheral blood (PB) cells (106 leukocytes per tube) were incubated with various combinations of mAb (Supplementary Table S3) for 15 minutes.

Manufacturer protocol Publication protocol 1 Relevant paper

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