PE Rat Anti-Mouse CD184

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a myocyte-depleted cardiac cell population was prepared, incubating minced myocardium in 0.1% collagenase IV (Gibco BrL) 45 min at 37 °C, lethal to most adult mouse cardiomyocytes (Zhou et al., 2000). Cells were then filtered through a 70-μm mesh. To exclude spurious effects of enzymatic digestion, BM cells with or without collagenase treatment were stained revealing no significantly changed staining of labeled cell antigens (data not shown). Cells were stained.		Cells were subjected to flow cytometry using EPICS XLMCL flow cytometer and Expo32 ADC Xa software (Beckman Coulter). Each analysis included 50,000 events.

Upstream tips
a myocyte-depleted cardiac cell population was prepared, incubating minced myocardium in 0.1% collagenase IV (Gibco BrL) 45 min at 37 °C, lethal to most adult mouse cardiomyocytes (Zhou et al., 2000). Cells were then filtered through a 70-μm mesh. To exclude spurious effects of enzymatic digestion, BM cells with or without collagenase treatment were stained revealing no significantly changed staining of labeled cell antigens (data not shown). Cells were stained.
Downstream tips
Cells were subjected to flow cytometry using EPICS XLMCL flow cytometer and Expo32 ADC Xa software (Beckman Coulter). Each analysis included 50,000 events.

Publication protocol

"8 to 10 weeks old C57BL/6 mice (n = 8) were treated with saline, G-CSF, Diprotin A, AMD3100 or combined daily for 6 days. Cells were separated from peripheral blood as described previously (Deindl et al., 2006). The following monoclonal antibodies were used: CD45-PerCP, CD34-FITC, CXCR4-PE (all from BD Pharmingen). Matching isotype antibodies (BD Pharmingen) served as controls. Cells were analyzed by 3-color flow cytometry using a Coulter® Epics® XL-MCLTM flow cytometer (Beckman Coulter). Each analysis included 50,000 events.

Cardiac cells of infarcted hearts of C57BL/6 mice were analyzed 6 days after MI (n = 8). Therefore, a myocyte-depleted cardiac cell population was prepared, incubating minced myocardium in 0.1% collagenase IV (Gibco BrL) 45 min at 37 °C, lethal to most adult mouse cardiomyocytes (Zhou et al., 2000). Cells were then filtered through a 70-μm mesh. To exclude spurious effects of enzymatic digestion, BM cells with or without collagenase treatment were stained revealing no significantly changed staining of labeled cell antigens (data not shown). Cells were stained with CD45-PerCP, CD34-FITC, c-kit-PE, CXCR4-PE, Sca1-PE, CD3-biotin, CD45R/B220-biotin, CD11b-biotin, TER-119-biotin, Ly-6G-biotin Abs (all from BD Pharmingen) and subjected to flow cytometry using EPICS XLMCL flow cytometer and Expo32 ADC Xa software (Beckman Coulter). Each analysis included 50,000 events."

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