Flow cytometry Anti-bodies Human - CD23

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).

Start discussion

No discussions found

Share your thoughts or question with experts in your field

Start a discussion

Found 3 matching solutions for this experiment

Anti-Human FMC7 FITC/CD23 PE/CD19 PerCP-Cy™5.5

BD Biosciences

Upstream tips	Protocol tips	Downstream tips
	The lymph node samples were cut, mechanically disintegrated, and then filtered. A specimen of 25–50 µL (<20 × 109 cells/L) was pipetted into five tubes and labelled with the Ab. Following a 15‐minutes incubation, red blood cells were automatically lysed and washed with Cell Wash solution (BD Biosciences) using BD FACS Lyse/Wash Assistant (BD Biosciences), followed by the automatic fixation of the specimen with Cell FIX solution (BD Biosciences).	Flow cytometry analysis of the specimens was performed with Becton Dickinson FACSCanto II. At least 20,000 cells were acquired for analysis.

Protocol tips
The lymph node samples were cut, mechanically disintegrated, and then filtered. A specimen of 25–50 µL (<20 × 109 cells/L) was pipetted into five tubes and labelled with the Ab. Following a 15‐minutes incubation, red blood cells were automatically lysed and washed with Cell Wash solution (BD Biosciences) using BD FACS Lyse/Wash Assistant (BD Biosciences), followed by the automatic fixation of the specimen with Cell FIX solution (BD Biosciences).
Downstream tips
Flow cytometry analysis of the specimens was performed with Becton Dickinson FACSCanto II. At least 20,000 cells were acquired for analysis.

Manufacturer protocol Publication protocol 1 Relevant paper

PE Mouse Anti-Human CD23

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. Then, erythrocytes were lysed in FACS-Lysing-Solution (BD Biosciences, San José, CA, USA). Washed cells were acquired on a FACSCalibur (BD Biosciences) and analyzed by FlowJo software (TreeStar, Ashland, OR, USA)

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. Then, erythrocytes were lysed in FACS-Lysing-Solution (BD Biosciences, San José, CA, USA). Washed cells were acquired on a FACSCalibur (BD Biosciences) and analyzed by FlowJo software (TreeStar, Ashland, OR, USA)

Manufacturer protocol Publication protocol 1 Relevant paper

CD23 Monoclonal Antibody (EBVCS2), eBioscience™

eBioscience

Upstream tips	Protocol tips	Downstream tips
Aliquots of 1.5 × 106 cells were used for each staining.	Before staining, cells were blocked with 10% vol/vol mouse serum (Life Technologies, Carlsbad, Calif).	Dead cells were excluded from the analysis with eFluor 780 Fixable Viability Dye (eBioscience). Flow cytometric analysis was performed on a Beckman Coulter FC 500 flow cytometer (Beckman Coulter).

Upstream tips
Aliquots of 1.5 × 106 cells were used for each staining.
Protocol tips
Before staining, cells were blocked with 10% vol/vol mouse serum (Life Technologies, Carlsbad, Calif).
Downstream tips
Dead cells were excluded from the analysis with eFluor 780 Fixable Viability Dye (eBioscience). Flow cytometric analysis was performed on a Beckman Coulter FC 500 flow cytometer (Beckman Coulter).

Manufacturer protocol Publication protocol 1 Relevant paper

Can't find the product you've used to perform this experiment? It would be great if you can help us by Adding a product!

Fill out your contact details and receive price quotes in your Inbox