PE Rat anti-Mouse CD34

Protocol tips

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	Non-specific binding was blocked by incubation in FACS buffer (Life Technologies) containing 0.5% BSA and 40 nM EDTA, (Sigma-Aldrich, Denmark). Following incubation, the unbound antibodies were removed using a 3 ml wash of FACS buffer.

Protocol tips
Non-specific binding was blocked by incubation in FACS buffer (Life Technologies) containing 0.5% BSA and 40 nM EDTA, (Sigma-Aldrich, Denmark). Following incubation, the unbound antibodies were removed using a 3 ml wash of FACS buffer.

Publication protocol

mBMSCs in cell suspension washed with PBS−/− (Life Technologies). Cells were stained with pre-conjugated CD34-PE antibody for 20 min on ice. Non-specific binding was blocked by incubation in FACS buffer (Life Technologies) containing 0.5% BSA and 40 nM EDTA, (Sigma-Aldrich, Denmark). Following incubation, the unbound antibodies were removed using a 3 ml wash of FACS buffer. The cells were sorted into 2 separate populations (CD34− and CD34+) using a FACS Aria™ III (BD Biosciences, Denmark). Sorting gates were established based on the appropriate isotype control for CD34 antibody (supplementary Table 2). The sorted cell populations were then re-cultured for further experiments.

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