Freshly dissected murine (C57BL/6, 10- to 12-week-old females) femurs and tibias (Charles River Laboratories) were flushed ×5 with 10 mL RPMI 1640 Medium (Thermo 11875093) + 10% FBS (Thermo 10082147). Cells were centrifuged at 300 ×g for 5 mins, resuspended in RPMI + 10% FBS, 40 μm filtered (Fisher 11587522), and centrifuged at 300 ×g for 5 mins. Supernatant was discarded and the pellet was resuspended in 2 mL ACK Lysing Buffer (Thermo A1049201) for 5 mins at room temperature. Monocytes were washed in PBS, centrifuged at 300 ×g for 5 mins, resuspended in 1 mL Recovery™ Cell Culture Freezing Medium (Thermo 12648010), and stored in liquid nitrogen until use. Bone marrow-derived macrophages were expanded and activated in RPMI + 10% FBS + 25% L929-cell conditioned media (LCCM) before experiments. IF staining confirmed that the cells were positive for intestinal macrophages markers such as CD45 (30-F11, BioLegend), CD68 (FA-11, BioLegend), CD11b (M1/70, BioLegend), F4/80 (BM8, BioLegend), and CX3CR1 (SA011F11, BioLegend) in both 2D and 3D cultures.
Tibias
Manufactured by Charles River Laboratories
The Tibias is a laboratory equipment designed for the preparation and analysis of bone samples. It is used to extract and process tibial bones from small animals, such as rodents, for research purposes. The core function of the Tibias is to facilitate the collection and preparation of tibial bone specimens for various scientific investigations, including but not limited to studies on bone biology, biomechanics, and pathology.
Automatically generated - may contain errors
Lab products found in correlation
Ack lysing buffer, by Thermo Fisher Scientific (4 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (4 mentions)
Recovery cell culture freezing medium, by Thermo Fisher Scientific (2 mentions)
Sa011f11, by BioLegend (2 mentions)
Fa 11, by BioLegend (2 mentions)
M1 70, by BioLegend (2 mentions)
C57bl 6, by Charles River Laboratories (2 mentions)
2 protocols using tibias
1
Isolation and Characterization of Murine Bone Marrow-Derived Macrophages
2
Isolation and Characterization of Murine Bone Marrow-Derived Macrophages
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Freshly dissected murine (C57BL/6, 10- to 12-week-old females) femurs and tibias (Charles River Laboratories) were flushed ×5 with 10 mL RPMI 1640 Medium (Thermo 11875093) + 10% FBS (Thermo 10082147). Cells were centrifuged at 300 ×g for 5 mins, resuspended in RPMI + 10% FBS, 40 μm filtered (Fisher 11587522), and centrifuged at 300 ×g for 5 mins. Supernatant was discarded and the pellet was resuspended in 2 mL ACK Lysing Buffer (Thermo A1049201) for 5 mins at room temperature. Monocytes were washed in PBS, centrifuged at 300 ×g for 5 mins, resuspended in 1 mL Recovery™ Cell Culture Freezing Medium (Thermo 12648010), and stored in liquid nitrogen until use. Bone marrow-derived macrophages were expanded and activated in RPMI + 10% FBS + 25% L929-cell conditioned media (LCCM) before experiments. IF staining confirmed that the cells were positive for intestinal macrophages markers such as CD45 (30-F11, BioLegend), CD68 (FA-11, BioLegend), CD11b (M1/70, BioLegend), F4/80 (BM8, BioLegend), and CX3CR1 (SA011F11, BioLegend) in both 2D and 3D cultures.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!