Take 20–30 μL of peripheral blood through the tail vein of the mouse and add it to the anticoagulation tube. Take the bone marrow cells from the femur and tibia of the sacrificed mice. The red blood cells were lysed, and the bone marrow cells were filtered using a 100-mm cell strainer. Monoclonal antibodies to Mac-1 (M1/70, Biolegend), Gr-1 (RB6-8C5, Biolegend), c-Kit (2B8, Biolegend), Lin mix (Gr1, CD4, CD3, CD8a, Ter119, B220, IgM) (Biolegend), CD34 (MEC14.7, Biolegend), Sca1 (D7, Biolegend), FcgRII/III (93, Biolegend), IL-7Ra (A7R34, Biolegend) (all used 50 ng per million cells) were used where indicated. After incubation with antibodies, the samples were analyzed using the Attune NxT flow cytometer (Thermo), and the results were analyzed using FlowJo software. Here, 7-aminoactinomycin D (7-AAD) (A1310, Life Technologies) was used to exclude dead cells.
Cd34 mec14
Manufactured by BioLegend
Sourced in United States
CD34 (MEC14.7) is a cell surface glycoprotein that is expressed on hematopoietic stem and progenitor cells. It functions as a cell-cell adhesion factor and plays a role in the regulation of early hematopoiesis.
Automatically generated - may contain errors
Lab products found in correlation
C kit 2b8, by BioLegend (3 mentions)
Sca1 d7, by BioLegend (3 mentions)
Mac 1 m1 70, by BioLegend (2 mentions)
Gr 1 rb6 8c5, by BioLegend (2 mentions)
Il 7ra a7r34, by BioLegend (2 mentions)
7 aminoactinomycin d 7 aad, by Thermo Fisher Scientific (2 mentions)
Attune nxt flow cytometer, by Thermo Fisher Scientific (2 mentions)
Sca 1 d7, by Thermo Fisher Scientific (1 mentions)
Ter119 ly 76, by BD (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Cd11b m1 70, by BD (1 mentions)
Cd45 30 f11, by BD (1 mentions)
Cd31 mec 13, by BD (1 mentions)
Cd44 im7, by Thermo Fisher Scientific (1 mentions)
Cd86 2331 fun 1, by BD (1 mentions)
Cd80 16 10a1, by BD (1 mentions)
Hmβ1 1, by BioLegend (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
4 protocols using cd34 mec14
1
Immunophenotyping Murine Hematopoietic Cells
2
Multicolor Flow Cytometry Panel
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CD11c (N418), CD19 (1D3), CD24 (M1/69), CD29 (HMb1-1), CD3ε (500A2), F4/80 (BM8), and Sca-1 (D7) were purchased from eBioscience, BrdU (B44), CD45 (30-F11), MCP-1 (2H5) and Ter119 (Ly-76) were purchased from BD Bioscience, CD206 (MMR), CD31 (390), and CD34 (MEC14.7) were purchased from BioLegend, and CD11b (M1/70.15) was purchased from Caltag.
3
Isolation and Characterization of Murine Hematopoietic Cells
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Take 20–30 μl of peripheral blood through the tail vein of the mouse and add to the anticoagulation tube. Take the bone marrow cells from the femur and tibia of the sacrificed mice. The red blood cells were lysed, and the bone marrow cells were filtered using a 100-μm cell strainer. Monoclonal antibodies to Mac-1 (M1/70, Biolegend), Gr-1 (RB6-8C5, Biolegend), c-Kit (2B8, Biolegend), Lin mix (Gr1, CD4, CD3, CD8a, Ter119, B220, IgM) (Biolegend), CD34 (MEC14.7, Biolegend), Sca1 (D7, Biolegend), FcγRII/III (93, Biolegend), IL-7Ra (A7R34, Biolegend) (all used as 50 ng per million cells) were used where indicated. After incubation with antibodies, the samples were analyzed using the Attune NxT flow cytometer (Thermo), and the results were analyzed using FlowJo software. Here, 7-aminoactinomycin D (7-AAD) (A1310, Life Technologies) was used to exclude dead cells.
4
Isolation and Characterization of Mouse Bone Marrow Mesenchymal Stem Cells
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Six- to eight-week-old DBA1J mouse bone marrow cells were collected by flushing femurs and tibias with Dulbecco’s modified Eagle’s medium (Gibco, Carlsbad, CA, USA) containing 2 mM L-glutamine (Gibco), 1% antibiotics (penicillin (10 U/ml)–streptomycin (10 g/ml)) (Gibco) and 15% heat-inactivated fetal bovine serum (FBS, Gibco) with an endotoxin level ≤5 EU/ml and hemoglobin level ≤10 mg/dl (Gibco)52 (link). When cells reached around 80% confluency, the medium was aspirated and 3–5 ml trypsin-EDTA (Gibco) were added to each dish. The dishes were then incubated for ~5 min to allow cell detachment. Next, an equal volume of culture medium was added to inactivate trypsin. The marrow cell immunophenotypes were persistently positive for Sca-1 (D7; BioLegend, San Diego, CA, USA), CD44 (IM7; eBioscience, San Diego, Ca, USA), and CD29 (HMβ1-1; BioLegend), but negative for c-Kit (2B8; BioLegend), CD11b (M1/70; BD Pharmingen, San Diego, CA, USA), CD34 (MEC14.7; BioLegend), CD106 (429 (MVCAM.A); BD Pharmingen), CD45 (30-F11; BD Pharmingen), CD31 (MEC 13.3; BD Pharmingen), CD80 (16-10A1, BD Pharmingen), and CD86 (2331 (FUN-1), BD Pharmingen) after more than 10 passages (two months of culturing) (Supplementary Fig. 1 ).
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