Flow cytometry Anti-bodies Mouse - CD252/OX40L

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

Biotin Rat Anti-Mouse OX40 Ligand (CD252)

BD Biosciences

Upstream tips	Protocol tips	Downstream tips
). Briefly, mouse BM was obtained from the long bones of the hind legs. After erythrocyte lysis, BM cells were suspended at 1×106/ml in RPMI 1640 with 10% FBS, 10 ng/ml murine GM-CSF and 1 ng/ml murine IL-4. Cells were plated into wells of six-well tissue culture plates, and cultured for 6 days;	All Abs used for DC FACS analysis were obtained from BD Pharmingen. After washing, DCs were resuspended in PBS containing 1% BSA and 0.1% NaN3, fixed with 1% paraformaldehyde, and analyzed by a FACScan flow cytometer (BD Immunocytometry Systems, Mountain View, CA)

Upstream tips
). Briefly, mouse BM was obtained from the long bones of the hind legs. After erythrocyte lysis, BM cells were suspended at 1×106/ml in RPMI 1640 with 10% FBS, 10 ng/ml murine GM-CSF and 1 ng/ml murine IL-4. Cells were plated into wells of six-well tissue culture plates, and cultured for 6 days;
Protocol tips
All Abs used for DC FACS analysis were obtained from BD Pharmingen. After washing, DCs were resuspended in PBS containing 1% BSA and 0.1% NaN3, fixed with 1% paraformaldehyde, and analyzed by a FACScan flow cytometer (BD Immunocytometry Systems, Mountain View, CA)

Manufacturer protocol Publication protocol 1 Relevant paper

PE/Cyanine7 anti-mouse CD252 (OX40L) Antibody

BioLegend

Upstream tips	Protocol tips	Downstream tips
single-cell suspensions from fresh mouse CLNs or cultured BM-macrophages were filtered by a 70-μm cell strainer, centrifuged, and resuspended. Cells were incubated with a stain kit (LIVE/DEAD Fixable Yellow Dead Cell Stain Kit, L34959, Thermo Fisher Scientific, Waltham, MA)

Upstream tips
single-cell suspensions from fresh mouse CLNs or cultured BM-macrophages were filtered by a 70-μm cell strainer, centrifuged, and resuspended. Cells were incubated with a stain kit (LIVE/DEAD Fixable Yellow Dead Cell Stain Kit, L34959, Thermo Fisher Scientific, Waltham, MA)

Manufacturer protocol Publication protocol 1 Relevant paper

APC anti-mouse CD252 (OX40L) Antibody

BioLegend

Upstream tips	Protocol tips	Downstream tips
		Data acquisition was performed using a flow cytometer (FACSCalibur, BD-Pharmingen, San Diego, CA, United States) by collecting a minimum of 10000 events per sample.

Downstream tips
Data acquisition was performed using a flow cytometer (FACSCalibur, BD-Pharmingen, San Diego, CA, United States) by collecting a minimum of 10000 events per sample.

Manufacturer protocol Publication protocol 1 Relevant paper

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