Pe cy7 conjugated anti human cd45 antibody

Newborn Ube3a^mat−/pat+ IL2rg^−/y mice (2–5 days old) were irradiated with 150 rads from a Cesium source and transplanted intrahepatically with 500 000 total human CD34+ HSPC/mouse. Eight weeks post-transplant, mice were bled via the tail vein and analyzed for engraftment by flow cytometry using a PE-CY7-conjugated anti-human CD45 antibody (BD Biosciences, San Jose, CA). Successfully engrafted mice were then evaluated for behavioral phenotypes. The order and age of testing were as follows: (1) Open field at 9 weeks of age, (2) Beam walking at 9 weeks of age, (3) Rotarod at 10 weeks of age, and (4) NOR at 11 weeks of age, (5) EEG at 15–17 weeks.
At 4–5 weeks of age, Ube3a^mat−/pat+ IL2rg^−/y mice were treated with 20 mg/kg busulfan intraperitonially 48 and 24 h prior to transplanting with 500 000 total human CD34+ HSPC/mouse intravenously. Six weeks post-transplant, mice were bled via the tail vein and analyzed for engraftment by flow cytometry using a PE-CY7-conjugated anti-human CD45 antibody (BD Biosciences, San Jose, CA). Successfully engrafted mice were then evaluated for behavioral phenotypes. The order and age of testing were as follows: (1) Open field at 11–12 weeks of age, (2) Beam walking at 11–12 weeks of age, (3) Rotarod at 12–13 weeks of age, (4) NOR at 14–15 weeks of age.

NOD-RAG1^−/− IL2rg^−/y (NRG) mice (stock number 007799) were obtained from The Jackson Laboratory (Sacramento, CA). Two to five-day old NRG mice (N = 15 per cohort) were sublethally irradiated with 100 rads from a Cesium source and transplanted intrahepatically with either the NT, EGFP control vector transduced, or the hAS8 vector transduced human CD34+ HSPC (300 000 total cells). To determine engraftment levels, at 12 weeks post-transplant, mice were bled via the tail vein and were analyzed by flow cytometry with a PE-CY7-conjugated anti-human CD45 antibody (BD Biosciences, San Jose, CA). Flow cytometry was performed on a Beckman Coulter FC-500 and analyzed by CXP software. Mice were used in accordance with institutional and IACUC guidelines.