Flow cytometry Anti-bodies Human - CD36/CB38

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

FITC Mouse Anti-Human CD36

BD Biosciences

Upstream tips	Protocol tips	Downstream tips
To perform flow cytometry analysis and to sort BFU-E and CFU-E cells from primary human samples including BM, cord blood, and peripheral blood, we isolated CD45+ cells from mononuclear cells (MNC) derived by Ficoll density gradient separation, followed by positive selection using CD45 magnetic beads.	Ab or appropriate isotype controls (all from BD Biosciences)were incubated for 30 minutes in the dark. Cells were washed twice with 40 mL phosphate-buffered saline/0.5% bovine serum albumin, resuspended in 5 mL phosphate-buffered saline/0.5% bovine serum albumin, and stained with the viability marker 7-AAD on ice for 10 minutes in the dark.	The analysis was performed using BD FACSDiva. Sorting was performed using a MoFlo high-speed cell sorter (Beckman Coulter). The number of BFU-E or CFU-E cells in one million MNCs was quantitated.

Upstream tips
To perform flow cytometry analysis and to sort BFU-E and CFU-E cells from primary human samples including BM, cord blood, and peripheral blood, we isolated CD45+ cells from mononuclear cells (MNC) derived by Ficoll density gradient separation, followed by positive selection using CD45 magnetic beads.
Protocol tips
Ab or appropriate isotype controls (all from BD Biosciences)were incubated for 30 minutes in the dark. Cells were washed twice with 40 mL phosphate-buffered saline/0.5% bovine serum albumin, resuspended in 5 mL phosphate-buffered saline/0.5% bovine serum albumin, and stained with the viability marker 7-AAD on ice for 10 minutes in the dark.
Downstream tips
The analysis was performed using BD FACSDiva. Sorting was performed using a MoFlo high-speed cell sorter (Beckman Coulter). The number of BFU-E or CFU-E cells in one million MNCs was quantitated.

Manufacturer protocol Publication protocol 1 Relevant paper

CD36 Monoclonal Antibody (eBioNL07 (NL07)), PerCP-eFluor 710, eBioscience™

eBioscience

Upstream tips	Protocol tips	Downstream tips
	>20 × 105 disaggregated sphere cells were stained with a stem cell‐specific antibody panel and analysed on a BDTM LSR II flow cytometer. To a volume of 100 μL cell suspension, 5 μL antibody mix was added, mixed and incubated for 20 minutes at 4°C in the dark and washed twice with 250 μL DPBS or FACS buffer.

Protocol tips
>20 × 105 disaggregated sphere cells were stained with a stem cell‐specific antibody panel and analysed on a BDTM LSR II flow cytometer. To a volume of 100 μL cell suspension, 5 μL antibody mix was added, mixed and incubated for 20 minutes at 4°C in the dark and washed twice with 250 μL DPBS or FACS buffer.

Manufacturer protocol Publication protocol 1 Relevant paper

Purified Mouse Anti-Human CD36

BD Biosciences

Upstream tips	Protocol tips	Downstream tips
	Fluorescently labelled antibodies were validated for their sensitivity using monocyte derived macrophages

Protocol tips
Fluorescently labelled antibodies were validated for their sensitivity using monocyte derived macrophages

Manufacturer protocol Publication protocol 1 Relevant paper

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