Flow cytometry Anti-bodies Human - CD96

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).

3 Matching solutions
Start a discussion

Start discussion

No discussions found

Start your discussion

Share your thoughts or question with experts in your field

Start a discussion

Found 3 matching solutions for this experiment

PE anti-human CD96 (TACTILE) Antibody

BioLegend

Upstream tips	Protocol tips	Downstream tips
	Cells were stained with Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) (BD Pharmingen, San Diego, CA, USA) to detect the apoptosis state in accordance with the manufacturer specifications.	The cells (1 × 106) were collected and resuspended in 100 μl Cell Staining Buffer, the antibody was added and incubated at 4 °C for 15 min, washed again with cell staining buffer one time, and then subject to the apoptosis assay. To exclude non-viable cells, cells were stained with 7AAD before flow cytometry (BD Versa, San Diego, CA, USA) analysis for antigen detecting.

Protocol tips
Cells were stained with Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) (BD Pharmingen, San Diego, CA, USA) to detect the apoptosis state in accordance with the manufacturer specifications.
Downstream tips
The cells (1 × 106) were collected and resuspended in 100 μl Cell Staining Buffer, the antibody was added and incubated at 4 °C for 15 min, washed again with cell staining buffer one time, and then subject to the apoptosis assay. To exclude non-viable cells, cells were stained with 7AAD before flow cytometry (BD Versa, San Diego, CA, USA) analysis for antigen detecting.

Manufacturer protocol Publication protocol 1 Relevant paper

CD96 (TACTILE) Monoclonal Antibody (NK92.39), PE, eBioscience™

eBioscience

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) as reported

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) as reported

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) as reported

Manufacturer protocol Publication protocol 1 Relevant paper

PE Mouse anti-Human CD96

BD Biosciences

Upstream tips	Protocol tips	Downstream tips
A total of 2 to 5 mL bone marrow was extracted with EDTA anticoagulation from AML patients before chemotherapy. The marrow was treated by density gradient centrifugation in Ficoll-Paque PREMIUM to isolate mononuclear cells.

Upstream tips
A total of 2 to 5 mL bone marrow was extracted with EDTA anticoagulation from AML patients before chemotherapy. The marrow was treated by density gradient centrifugation in Ficoll-Paque PREMIUM to isolate mononuclear cells.

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!

Outsource your experiment

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

Outsource experiment