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Biotinylated lineage cocktail

Manufactured by BioLegend

Biotinylated-lineage cocktail is a laboratory product designed to facilitate cell identification and sorting. It contains a mixture of antibodies that are conjugated with biotin, a small molecule that can be detected by streptavidin-based detection systems. The core function of this product is to label specific cell types, allowing researchers to isolate and analyze them using flow cytometry or other cell sorting techniques.

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2 protocols using biotinylated lineage cocktail

1

Magnetic Isolation of Neutrophils and LSK+ Cells

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For magnetic isolation of blood neutrophils, red blood cells were lysed as above, and single cell suspensions were washed in selection media. Cells were then incubated with Fc block for 10 min on ice as noted above. Neutrophils were isolated using the MojoSort Mouse Ly6G Selection Kit (Biolegend, Cat# 480,058) according to manufacturer’s instructions. For magnetic isolation of femur bone marrow LSK + cells, single cell suspensions were blocked and stained with biotinylated-lineage cocktail (Biolegend, Cat# 133,307) and stem/progenitor cells were negatively selected using streptavidin-conjugated nanobeads (Biolegend, Cat# 480,015). Lineage-negative cells were then stained with c-Kit-APC (Biolegend, Cat# 105,811) for positive selection with anti-APC-conjugated nanobeads (Biolegend, Cat# 480,071). Finally, Lin-cKit + bone marrow cells were stained with Sca1-APC (Biolegend, Cat# 108,111) for positive selection with anti-APC-conjugated nanobeads. The resulting neutrophil and LSK + cell populations were stored in Qiazol lysis reagent (Qiagen, Cat# 79,306) at -80 °C.
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2

Magnetic Isolation of Neutrophils and LSK+ Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
For magnetic isolation of blood neutrophils, red blood cells were lysed as above, and single cell suspensions were washed in selection media. Cells were then incubated with Fc block for 10 min on ice as noted above. Neutrophils were isolated using the MojoSort Mouse Ly6G Selection Kit (Biolegend, Cat# 480058) according to manufacturer’s instructions. For magnetic isolation of femur bone marrow LSK+ cells, single cell suspensions were blocked and stained with biotinylated-lineage cocktail (Biolegend, Cat# 133307) and stem/progenitor cells were negatively selected using streptavidin-conjugated nanobeads (Biolegend, Cat# 480015). Lineage-negative cells were then stained with c-Kit-APC (Biolegend, Cat# 105811) for positive selection with anti-APC-conjugated nanobeads (Biolegend, Cat# 480071). Finally, Lin-cKit+ bone marrow cells were stained with Sca1-APC (Biolegend, Cat# 108111) for positive selection with anti-APC-conjugated nanobeads. The resulting neutrophil and LSK+ cell populations were stored in Qiazol lysis reagent (Qiagen, Cat# 79306) at −80 °C.
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