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

Protocol tips

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.

Publication protocol

Flow cytometry was adopted to determine the percentages of CD40, CD80, CD204 and CD206 as macrophage surface antigens. 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 with FITC-labeled CD40 antibody, CD80 antibody, CD204 antibody and CD206 antibody 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 to detect CD40, CD80, CD204 and CD206 with BD FACSARIA II Flow Cytometer of Becton-Dickinson and Company (BD). The detection results were analyzed with Flowjo in percentage.

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