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The Mouse Lineage Panel consisting of biotinylated CD3, B220, CD11b, GR-1 and Ter119 were purchased from BD Pharmingen.
The cells were "lineage depleted" by labeling the cells with biotin-conjugated lineage panel antibodies against B220, CD3ε, Gr-1, Mac-1 and Ter-119 (BD/Pharmingen, San Diego, CA).
Lineage-negative (Linneg) cells were identified by gating against cells positive for the mouse lineage panel + CD31.
Bone marrow cells were collected from femurs and immunophenotyped using a mouse lineage panel as described elsewhere [ 3].
Murine HSPCs were purified by lin− selection, using the Biotin Mouse Lineage Panel (BD PharMingen) and the StemSep Mouse Progenitor Enrichment Kit (Stem Cell Technologies).
These antibodies included a biotin-conjugated anti-mouse hematopoietic lineage panel supplemented with anti-mouse CD31-biotin, anti-mouse CD133-phycoerythrin (CD-133-PE) and anti-mouse CD24-fluorescein isothiocyanate (CD24-FITC), each at a 1 100 dilution.
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The following monoclonal antibodies were used: anti-mouse lineage panel-APC, anti-c-Kit-RPE, anti-Sca-1-PE/Cy7, anti-CD127-Pacific Blue (all BD Biosciences), anti-CD34-FITC, anti-CD16/CD32-PE/Cy5 and biotinylated anti-CD135 (all eBioscience, Insight Biotechnology, UK).
In contrast, d 0.5) is the most powerful for detecting change in moderately abundant lineages (Panel 5).
d U shows an opposite trend: it is the most powerful for detecting change in rare lineages (Panel 6) but has almost no power for change in most abundant lineages (Panel 4).
For the evenness change scenario (Panel 2), the power of d W and d 0.5) is very close and is more powerful than d(0) and d U. d W is the most powerful for detecting change in most abundant lineages (Panel 4) but is much less powerful for change in rare lineages (Panel 6).
The matrix (M) gene of the 3 iswine/Guangdong/223/2006211/2006 (H3N2), swine/Guanddong/223/2006 (H3N2), and swine/Guangdong/423/2006 (H3N2)—grouped in classical swine lineage (Technical Appendix, panel I), and other internal genes were located in intermediate human subtype H3N2 lineage (Technical Appendix, panels E H and J).
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