All mouse experiments were approved by the Austrian Ministry of Education, Science and Research (BMBWF-66.010/0056-V/3b/2019). 5x105 CRISPR-modified HSPCs were intrafemorally transplanted into irradiated 8–12-week-old NSG mice. For competitive transplants, 1x105 sort-purified HSPCs (1:1 mixture of CALR INS and WT cells) were intrahepatically transplanted into new-born NSGW41 mice. Human engraftment in murine bone marrow was evaluated after 8, 16 and 24 weeks via flow cytometry. Cells were stained with mTer119-BUV661, mCD45-APC-Cy7, hCD45-BB700, CD33-PE (BD Biosciences), CD19-SB600 (eBioscience, San Diego, CA, USA), CD41-PE-Cy7 (BioLegend) antibodies and SYTOX Red (Invitrogen, Carlsbad, CA, USA) and measured on a CytoFLEX LX flow cytometer (Beckman Coulter).
Hcd45 bb700
Manufactured by BD
Sourced in United States
The HCD45-BB700 is a laboratory centrifuge designed for general-purpose applications. It features a maximum speed of 4,500 RPM and a maximum relative centrifugal force (RCF) of 2,700 g. The centrifuge has a rotor capacity of 4 x 700 mL.
Automatically generated - may contain errors
2 protocols using hcd45 bb700
1
Engraftment of CRISPR-modified HSPCs
2
Xenograft Model of HSPC Transplantation
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All mouse experiments were approved by the Austrian Ministry of Education, Science and Research (BMBWF-66.010/0056-V/3b/2019). 5 × 105 CRISPR-modified HSPCs were intrafemorally transplanted into irradiated 8–12-week-old NSG mice. For competitive transplants, 1 × 105 sort-purified HSPCs (1:1 mixture of CALR INS and WT cells) were intrahepatically transplanted into new-born NSGW41 mice. Human engraftment in murine bone marrow was evaluated after 8, 16, and 24 weeks via flow cytometry. Cells were stained with mTer119-BUV661, mCD45-APC-Cy7, hCD45-BB700, CD33-PE (BD Biosciences), CD19-SB600 (eBioscience, San Diego, CA, USA), CD41-PE-Cy7 (BioLegend) antibodies and SYTOX Red (Invitrogen, Carlsbad, CA, USA) and measured on a CytoFLEX LX flow cytometer (Beckman Coulter).
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