Single cell suspensions were prepared by cutting lungs into small fragments followed by enzymatic digestion. For this, 1 ml IMDMc with 100U/ml collagenase type IV (Sigma-Aldrich) was added per lung lobe for 60 min incubation at 37 °C. Cold IMDMc containing 10 mM EDTA and 20 mM HEPES pH 7.5 was then added, and cells were filtered though 70 µm nylon sieves (BD Biosciences). Red blood cells were lysed by hypotonic lysis. Fc-receptors were blocked with anti-CD16/CD32 for 30 min, followed by staining DC- and macrophage-subsets with a cocktail of mAbs specific to CD11c, CD11b and MHC class II (I-A/I-E) and the different Notch ligands (for clone identifiers see at list of mAbs above).
70 m nylon sieves
Manufactured by BD
The 70 µm nylon sieves are laboratory equipment designed to separate particles based on size. They feature a nylon mesh with a pore size of 70 micrometers, allowing the passage of smaller particles while retaining larger ones.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using 70 m nylon sieves
1
Isolation and Characterization of Lung Immune Cells
2
Isolation and Characterization of Lung Immune Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Single cell suspensions were prepared by cutting lungs into small fragments followed by enzymatic digestion. For this, 1 ml IMDMc with 100U/ml collagenase type IV (Sigma-Aldrich) was added per lung lobe for 60 min incubation at 37 °C. Cold IMDMc containing 10 mM EDTA and 20 mM HEPES pH 7.5 was then added, and cells were filtered though 70 µm nylon sieves (BD Biosciences). Red blood cells were lysed by hypotonic lysis. Fc-receptors were blocked with anti-CD16/CD32 for 30 min, followed by staining DC- and macrophage-subsets with a cocktail of mAbs specific to CD11c, CD11b and MHC class II (I-A/I-E) and the different Notch ligands (for clone identifiers see at list of mAbs above).
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!