PE Mouse Anti-Human CD44

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	PC-3 cells and PC/DX25 cells (1 × 106) were resuspended in 100 μL of staining buffer. Fluorescent-conjugated antibodies were added and cells were dissociated, antibody-labeled (1:100 dilution per 106 cells), incubated for 30 min on ice and resuspended in Hanks’ Balanced Salt Solution (HBSS; Invitrogen, Waltham, Massachusetts, USA) containing 2% FBS and 10mM HEPES (Invitrogen).

Protocol tips
PC-3 cells and PC/DX25 cells (1 × 106) were resuspended in 100 μL of staining buffer. Fluorescent-conjugated antibodies were added and cells were dissociated, antibody-labeled (1:100 dilution per 106 cells), incubated for 30 min on ice and resuspended in Hanks’ Balanced Salt Solution (HBSS; Invitrogen, Waltham, Massachusetts, USA) containing 2% FBS and 10mM HEPES (Invitrogen).

Publication protocol

For surface maker analysis, PC-3 cells and PC/DX25 cells (1 × 106) were resuspended in 100 μL of staining buffer. Fluorescent-conjugated antibodies were added and cells were dissociated, antibody-labeled (1:100 dilution per 106 cells), incubated for 30 min on ice and resuspended in Hanks’ Balanced Salt Solution (HBSS; Invitrogen, Waltham, Massachusetts, USA) containing 2% FBS and 10mM HEPES (Invitrogen). The PE-conjugated anti-CD44 antibody (BD Biosciences, San Jose, CA, USA) was used. Flow cytometry was done using a FACSCaliburTM flow cytometer (BD Biosciences). FlowJo software (Ver10.0, (BD) Becton, Dickinson and Company, Franklin Lakes, New Jersey, USA) was used to analyze the data.

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