Flow cytometry Anti-bodies Human - CD163

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

CD163 Antibody, anti-human, PE-Vio® 770, REAfinity™

Miltenyibiotec

Upstream tips	Protocol tips	Downstream tips
	A total of 0.1 ml of peripheral blood was incubated with 10 μl of antibody for 15 min at room temperature, then erythrocytes were lysed and leucocytes post-fixed. Afterwards, the flow cytometry analysis was performed.	Flow cytometric analysis was performed using a Navios Flow Cytometer and the Kaluza analysis software (Beckman Coulter, Milan, Italy), evaluating a total of 5 × 106 cells and detecting more than 30 events in the smallest subset investigated, according to consensus guidelines on the minimal residual disease

Protocol tips
A total of 0.1 ml of peripheral blood was incubated with 10 μl of antibody for 15 min at room temperature, then erythrocytes were lysed and leucocytes post-fixed. Afterwards, the flow cytometry analysis was performed.
Downstream tips
Flow cytometric analysis was performed using a Navios Flow Cytometer and the Kaluza analysis software (Beckman Coulter, Milan, Italy), evaluating a total of 5 × 106 cells and detecting more than 30 events in the smallest subset investigated, according to consensus guidelines on the minimal residual disease

Manufacturer protocol Publication protocol 1 Relevant paper

PE Mouse Anti-Human CD163

BD Biosciences

Upstream tips	Protocol tips	Downstream tips
	Cells were then detached with accutase (Sigma-Aldrich), washed in PBS, and incubated with 100 µL of blocking buffer [PBS containing 10% human AB serum (Sigma-Aldrich), 2% FCS (Lonza), and 0.02% NaN3 (Sigma-Aldrich)]. Cells were then labeled in brilliant stain buffer (BD Bioscience) with a combination of fluorescently conjugated monoclonal antibodies	Flow cytometry analysis was performed on a BD LSRFortessa instrument using FACSDiva software (BD Biosciences), with 10,000 events acquired for each sample.

Protocol tips
Cells were then detached with accutase (Sigma-Aldrich), washed in PBS, and incubated with 100 µL of blocking buffer [PBS containing 10% human AB serum (Sigma-Aldrich), 2% FCS (Lonza), and 0.02% NaN3 (Sigma-Aldrich)]. Cells were then labeled in brilliant stain buffer (BD Bioscience) with a combination of fluorescently conjugated monoclonal antibodies
Downstream tips
Flow cytometry analysis was performed on a BD LSRFortessa instrument using FACSDiva software (BD Biosciences), with 10,000 events acquired for each sample.

Manufacturer protocol Publication protocol 1 Relevant paper

CD163 Monoclonal Antibody (eBioGHI/61 (GHI/61)), eBioscience™

eBioscience

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

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

Manufacturer protocol Publication protocol 1 Relevant paper

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