Toll-Like Receptor 4 Knockout (TLR4 KO) Mouse Bone Marrow-Derived Endothelial Cells from Creative Bioarray are isolated from bone marrow tissue of pathogen-free laboratory mice. Toll-Like Receptor 4 Knockout (TLR4 KO) Mouse Bone Marrow-Derived Endothelial Cells are grown in T25 tissue culture flasks pre-coated with gelatin-based coating solution for 2 min and incubated in Creative Bioarray’Culture Complete Growth Medium generally for 3-7 days. Cultures are then expanded. Prior to shipping, cells are detached from flasks and immediately cryo-preserved in vials. Each vial contains at least 1x10^6cells per ml and are delivered frozen. The method we use to isolate endothelial cells was developed based on a combination of established and our proprietary methods. These cells are pre-coated with PECAM-1 antibody, following the application of magnetic pre-coated with secondary antibody.
Toll-Like Receptor 4 Knockout (TLR4 KO) Mouse Bone Marrow-Derived Endothelial Cells are tested for expression of markers using antibody, VE-cadherin (CD144, VE-cadherin Antibody, C-19, sc6458, Santa Cruz); AF1002 (R&D System) or CD31/PECAM-1 (Purified Rat Anti-Mouse CD31, Catalog No. 553370, BD) by immunofluorescence staining or FACS. Toll-Like Receptor 4 Knockout (TLR4 KO) Mouse Bone Marrow-Derived Endothelial Cells are negative for bacteria, yeast, fungi and mycoplasma. Cells can be expanded for 3-6 passages at a split ratio of 1:2 under the cell culture conditions specified byCreative Bioarray. Repeated freezing and thawing of cells is not recommended.
Storage and Shipping
Creative Bioarray ships frozen cells on dry ice. On receipt, immediately transfer frozen cells to liquid nitrogen (-180 °C) until ready for experimental use. Live cell shipment is also available on request.
Never can primary cells be kept at -20 °C.
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