The BM was flushed from the tibia and femur of each mouse. Red blood cells (RBCs) were lysed once for 5 min at 4°C in 0.15 M NH4Cl (StemCell Technologies), washed once with PBS (Gibco), and counted using a cell counter (LUNATM Automated Cell Counter). For the detection of LSK cells and LT-HSCs, Lineage+ cells were removed by magnetic depletion using biotinylated lineage-specific antibodies (CD5, CD45R, CD11b, Gr-1, and Ter-119), followed by depletion with MACs beads conjugated to a monoclonal anti-biotin (Miltenyi Biotec). Lineage− cells were stained with phycoerythrin PE-Cy7-conjugated antibodies to Sca1 (558162), APC-conjugated antibodies to c-kit (553356), FITC-conjugated antibodies to CD48 (557484), and PE-conjugated antibodies to CD150 (561540), all from BD Biosciences. Cells were further stained with streptavidin-pacific blue (PB) (Invitrogen, S11222). Data were collected on a BD AriaIII (BD Biosciences) and analyzed using the FlowJo software (Tree Star).
Monoclonal anti biotin
Manufactured by Miltenyi Biotec
Monoclonal anti-biotin is a laboratory reagent used to detect and isolate biotinylated molecules. It binds specifically to the biotin moiety, allowing for the identification and purification of biotinylated proteins, nucleic acids, and other biomolecules.
Automatically generated - may contain errors
Lab products found in correlation
Aria 3, by BD (2 mentions)
Macs bead, by Miltenyi Biotec (2 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (2 mentions)
Streptavidin pacific blue, by Thermo Fisher Scientific (2 mentions)
0.15 m nh4cl, by STEMCELL (2 mentions)
Fitc conjugated antibodies to cd48 557484, by BD (1 mentions)
Pe conjugated antibodies to cd150 561540, by BD (1 mentions)
Lsrii system, by BD (1 mentions)
2 protocols using monoclonal anti biotin
1
Multiparameter Characterization of Murine HSCs
2
Isolation and Characterization of HSCs, MSCs, and Osteoblasts
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Example 4
BM was isolated from the tibiae and femurs of each mouse. Red blood cells (RBCs) were hemolyzed once for 5 min at 4° C. in 0.15 M NH4Cl (StemCell Technologies), washed once with PBS (Gibco), and counted using a hemocytometer. For detecting HSCs, MSCs, or osteoblasts, Lin+ cells were removed by using magnetic depletion using biotinylated lineage-specific antibodies (CD5, CD45R, CD11b, Gr-1, and Ter-119), followed by depletion with MACs beads conjugated to a monoclonal anti-biotin (Miltenyi Biotec). For staining of HSCs, Lin− cells were stained with phycoerythrin PE-Cy7-conjugated antibodies to Sca1 (558162), APC-conjugated antibodies to c-Kit (553356), FITC-conjugated antibodies to CD48 (557484), and PE-conjugated antibodies to CD150 (561540), all from Sciences. For MSCs and osteoblasts staining, Lin− cells were stained with APC-Cy7-CD45 (557659), APC-CD31 (551262), PE-Cy7-Sca1 (558162), or PE-CD51 (551187) (all from BD Sciences). Cells were further stained with streptavidin-pacific blue (PB) (Invitrogen, S11222). Data were collected on a BD LSRII system and AriaIII (BD Sciences) and analyzed using FlowJo software (Tress Star).
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