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MB Sample ID: SA093607
| Local Sample ID: | bovine_CL_LD_replicate1 |
| Subject ID: | SU001358 |
| Subject Type: | Mammal |
| Subject Species: | Bos taurus |
| Taxonomy ID: | 9913 |
| Gender: | Female |
| Animal Animal Supplier: | JBS Beef Plant 3435 Edward Babe Gomez Ave, Omaha, NE 68107 |
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Collection:
| Collection ID: | CO001352 |
| Collection Summary: | Tissue (~2.5 g) was washed thoroughly in TE buffer (10 mM Tris, 1 mM EDTA, pH 7.4). Minced tissue was resuspended in 10 mL tissue homogenate buffer (60% sucrose w/v in TE buffer containing protease and phosphatase inhibitor cocktails) and homogenized with a Teflon Dounce homogenizer in a glass vessel. The post-nuclear supernatant (PNS) fraction was obtained after centrifugation at 2000 rcf for 10 min. The supernatant was loaded into a 30 mL ultracentrifuge tube and overlaid sequentially with 40%, 25%, 10%, and 0% sucrose w/v in TE buffer containing protease and phosphatase inhibitor cocktails. Samples were centrifuged at 110,000 × g (ravg) for 30 min at 4 °C with no brake in a Beckman Coulter Avanti J-20 XP ultracentrifuge using an SW 32 Ti rotor. The LDs concentrated in a yellow-ish band at the top of the gradient were harvested and concentrated by centrifugation at 2000 rcf for 10 min at 4 °C. This protocol was derived from Ding et al. 2012, and Brasaemale et al. 2016. Ding, Y., Zhang, S., Yang, L., Na, H., Zhang, P., Zhang, H., … Liu, P. (2013). Isolating lipid droplets from multiple species. Nature Protocols, 8(1), 43–51. https://doi.org/10.1038/nprot.2012.142 Brasaemle, D. L., & Wolins, N. E. (2016). Isolation of Lipid Droplets from Cells by Density Gradient Centrifugation. Current Protocols in Cell Biology, 72, 3.15.1-3.15.13. https://doi.org/10.1002/cpcb.10 |
| Sample Type: | Ovary |
| Volumeoramount Collected: | 2.5 g of corpus luteum tissue |
