Total bone marrow was isolated from bones (tibia, femur, pelvis and vertebral column). Cells were then stained with anti–c‐Kit‐APC (BioLegend; 105812) antibody at 4°C for 30 min. c‐Kit+ cells from total bone marrow (BM) were enriched through magnetic‐activated cell separation (MACS; Miltenyi Biotec) according to the manufacturer’s protocol (LS Column, 130‐042‐402; anti–APC‐microbeads, 130‐090‐855). Cells were incubated with a lineage cocktail containing biotinylated antibodies against CD4 (BioLegend; 100508), CD8 (BioLegend; 100704), TER‐119 (BioLegend; 116204), CD11b (BioLegend; 101204), Gr‐1 (BioLegend; 108404), and B220 (BioLegend; 103222) at 4°C for 30 min. After washing, cells were incubated with anti–Sca‐1‐PE‐Cy7 (BioLegend; 108114) and anti–c‐Kit‐APC (BioLegend; 105812) antibodies. Biotinylated antibodies were developed with streptavidin‐APC‐Cy7 (BioLegend; 405208). LSK cells, which contain hematopoietic stem cells (HSCs) and multipotent progenitors (MPPs), were FACS sorted by selecting for Sca‐1+, c‐Kit+, lineage− cells.
Anti c kit apc
Manufactured by BioLegend
Anti–c‐Kit‐APC is a fluorescence-labeled antibody that binds to the c-Kit protein, also known as CD117. c-Kit is a receptor tyrosine kinase that plays a crucial role in the development and function of various cell types, including hematopoietic stem cells, mast cells, and melanocytes. The APC (Allophycocyanin) fluorophore is conjugated to the antibody, enabling detection and analysis of c-Kit-expressing cells using flow cytometry or other fluorescence-based applications.
Automatically generated - may contain errors
Lab products found in correlation
Anti sca1 pe cy7, by BioLegend (2 mentions)
Anti epcam apc, by Thermo Fisher Scientific (1 mentions)
Anti cd71 pe, by Thermo Fisher Scientific (1 mentions)
Facsaria sorp cell sorter, by BD (1 mentions)
Alexa fluor 647 donkey anti mouse igg, by Thermo Fisher Scientific (1 mentions)
Ter119, by BioLegend (1 mentions)
Streptavidin apc cy7, by BioLegend (1 mentions)
Magnetic activated cell sorting (macs), by Miltenyi Biotec (1 mentions)
Cd11b, by BioLegend (1 mentions)
Anti b220, by BioLegend (1 mentions)
Anti gr1, by BioLegend (1 mentions)
Anti sca 1 pe, by BioLegend (1 mentions)
Anti cd3, by BioLegend (1 mentions)
Anti ter119, by BioLegend (1 mentions)
Stemspan, by STEMCELL (1 mentions)
Ack lysis buffer, by Thermo Fisher Scientific (1 mentions)
Anti cd4, by BioLegend (1 mentions)
Anti cd11b, by BioLegend (1 mentions)
Anti cd8, by BioLegend (1 mentions)
Streptavidin pe, by BioLegend (1 mentions)
4 protocols using anti c kit apc
1
Isolation and Characterization of Murine Hematopoietic Stem/Progenitor Cells
2
Multiparametric Liver Cell Sorting
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Human or mouse liver single-cell suspensions were incubated with anti-DLK-FITC (MBL, D187-4, 1:100), anti-EpCAM-APC (eBioscience, 17-5791-82, 1:50), anti-NCAM1 (Sangon Biotech, D198946, 1:50), Alexa Fluor 647 donkey anti-mouse IgG (Invitrogen, A31571, 1:1000), anti-cKit-APC (BioLegend, 105812, 1:200), or anti-CD71-PE (eBioscience, 113808, 1:200) antibodies at 4 °C for 20 min. After washing once with PBS, cells were analyzed and sorted using a FACS Aria SORP cell sorter (BD Biosciences). Dead cells were excluded by DAPI staining.
3
Hematopoietic Stem Cell Maintenance in Hypoxic Conditions
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Upon crushing of long bones, hematopoietic cells were collected, filtered, and subjected to red blood cells lysis in ACK lysis buffer (Gibco); recovered cells were then washed and stained for fluorescence-activated cell sorting with anti-CD3, anti-CD8, anti-CD4, anti-B220, anti-CD11b, anti-Gr1, anti-Ter119 (all biotinylated), anti-SCA1-PE, and anti-C-KIT-APC (BioLegend). Freshly sorted HSC cells were seeded on a layer of confluent PαS+ cells in StemSpan (STEMCELL Technologies) supplemented with 50 ng TPO and 50 ng SCF at 37°C in 1% oxygen concentration. When cultured with conditioned media derived from PαS+ cells, HSC cells were maintained in 70% StemSpan supplemented with 50 ng TPO and 50 ng SCF fresh medium and 30% conditioned medium. All coculture experiments and all cultures with conditioned media from PαS+ cells have been performed in at least three independent experiments. Results have been normalized compared to control conditions and a pool of experiments is shown in the figures.
4
Isolation and Characterization of Murine Hematopoietic Progenitor Cells
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Experimental procedures were approved by the Dier Experimenten Commissie of the Royal Netherlands Academy of Arts and Sciences and performed according to the guidelines.
Primary bone marrow cells were harvested from 3-months-old C57BL/6 mice. Femur and Tibia were extracted, the bones ends were cut away to access the bone marrow which was flushed out using a 22G syringe with HBSS (-Ca, -Mg, -phenol red, Gibco 14175053) supplemented with Pen-Strep and 1% FCS. The bone marrow was dissociated and debris was removed by passing it through a 70 µm cell strainer (Corning, 431751). Cells were washed with 25 ml supplemented HBSS before linage marker staining was performed following the instructions of the EasySep™ Mouse Hematopoietic Progenitor Cell Isolation Kit (Stemcell) at half of the recommended concentration of the biotinylated antibodies. This was followed by 30min incubation at 4 o C with a Streptavidin-PE (Biolegend, 1:5000), anti c-kit-APC (Biolegend, 1:800) and anti sca1-PeCy7 (Biolegend, 1:400). After 2 additional washes with HBBS (+PS, +FCS) cells were prepared following the sortChIC protocol for the 4 different histone modifications.
Primary bone marrow cells were harvested from 3-months-old C57BL/6 mice. Femur and Tibia were extracted, the bones ends were cut away to access the bone marrow which was flushed out using a 22G syringe with HBSS (-Ca, -Mg, -phenol red, Gibco 14175053) supplemented with Pen-Strep and 1% FCS. The bone marrow was dissociated and debris was removed by passing it through a 70 µm cell strainer (Corning, 431751). Cells were washed with 25 ml supplemented HBSS before linage marker staining was performed following the instructions of the EasySep™ Mouse Hematopoietic Progenitor Cell Isolation Kit (Stemcell) at half of the recommended concentration of the biotinylated antibodies. This was followed by 30min incubation at 4 o C with a Streptavidin-PE (Biolegend, 1:5000), anti c-kit-APC (Biolegend, 1:800) and anti sca1-PeCy7 (Biolegend, 1:400). After 2 additional washes with HBBS (+PS, +FCS) cells were prepared following the sortChIC protocol for the 4 different histone modifications.
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