Flow cytometry Anti-bodies Mouse - CD204

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 2 matching solutions for this experiment

Recombinant Anti-CD204/MSR1 Antibody (FITC), Rabbit Monoclonal

Sino Biological Inc.

Upstream tips	Protocol tips	Downstream tips
	First, the four KC groups were cultured for 12 h and washed with PBS once to collect 1x106 adherent cells into flow cytometry tubes. Next, the collected cells were rinsed with 1-ml PBS twice and re-suspended with 200-µl PBS. Then KCs were incubated at 4˚C for 30 min. After washed 3 times with 1-ml PBS, the cells were centrifuged at 1,038 x g at 4˚C for 5 min, washed, and re-suspended with 200-µl PBS.	Detection with BD FACSARIA II Flow Cytometer of Becton-Dickinson and Company (BD). The detection results were analyzed with Flowjo in percentage.

Protocol tips
First, the four KC groups were cultured for 12 h and washed with PBS once to collect 1x106 adherent cells into flow cytometry tubes. Next, the collected cells were rinsed with 1-ml PBS twice and re-suspended with 200-µl PBS. Then KCs were incubated at 4˚C for 30 min. After washed 3 times with 1-ml PBS, the cells were centrifuged at 1,038 x g at 4˚C for 5 min, washed, and re-suspended with 200-µl PBS.
Downstream tips
Detection with BD FACSARIA II Flow Cytometer of Becton-Dickinson and Company (BD). The detection results were analyzed with Flowjo in percentage.

Manufacturer protocol Publication protocol 1 Relevant paper

CD204 antibody | 2F8

Bio-Rad Laboratories

Upstream tips	Protocol tips	Downstream tips
Single-cell suspensions were incubated with a Fc receptor block (1 μg/1 × 106 cells; BD Bioscience, San Diego, CA) to reduce nonspecific anti-body binding.	Dead cells were excluded using a Live/Dead Fixable Blue Dead Cell Stain Kit (Invitrogen) or 4,6-diamidino-2-phenylindole dihydrochloride (DAPI) (Sigma-Aldrich, St. Louis, MO).	Flow cytometry was performed using BD LSR II and BD FACS Aria III flow cytometers (BD Bioscience), and data were analyzed with FlowJo software (TreeStar, Ashland, OR).

Upstream tips
Single-cell suspensions were incubated with a Fc receptor block (1 μg/1 × 106 cells; BD Bioscience, San Diego, CA) to reduce nonspecific anti-body binding.
Protocol tips
Dead cells were excluded using a Live/Dead Fixable Blue Dead Cell Stain Kit (Invitrogen) or 4,6-diamidino-2-phenylindole dihydrochloride (DAPI) (Sigma-Aldrich, St. Louis, MO).
Downstream tips
Flow cytometry was performed using BD LSR II and BD FACS Aria III flow cytometers (BD Bioscience), and data were analyzed with FlowJo software (TreeStar, Ashland, OR).

Manufacturer protocol Publication protocol 1 Relevant paper

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