Tumors were minced, incubated in 100 units/mL collagenase IV (Thermo Fisher Scientific, Waltham, MA) and RPMI supplemented with 10% FBS for 30 minutes at 37°C in a Stomacher machine set at normal speed, washed through 40-micron nylon cell strainers (Falcon; BD Biosciences) in PBS with 2% FBS. After washing, cells were lysed using 1X BD Pharm Lyse Buffer (BD Biosciences). Cells were immediately stained and subsequently analyzed by flow cytometry (Live/Dead-PacOrange, Invitrogen; CD4-PE, H129.19, BD Biosciences; CD3-AF488, 145-2C11, BD Biosciences; CD8-PerCP-Cy5.5, 53–6.7, BD Biosciences; FOXP3-APC, FJK-16 s, eBioscience; EGFRvIII-specific CAR-AlexaFluor 647 multimer). Intracellular staining was performed for FOXP3 using FOXP3 Transcription Factor Staining Buffer Kit (ThermoFisher Scientific; Invitrogen).
40 micron nylon cell strainers
Manufactured by BD
Sourced in United States
The 40-micron nylon cell strainers are designed to separate cells and remove cellular debris from cell suspensions. They feature a nylon mesh with 40-micron pore size, allowing the passage of single cells while retaining larger particles.
Automatically generated - may contain errors
2 protocols using 40 micron nylon cell strainers
1
Tumor Cell Isolation and Analysis
2
Murine Splenocyte Isolation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Spleens were harvested from mice and mashed through 40-micron Nylon cell strainers (Falcon®, Corning, Inc., New York, NY, USA). Flow-through cells were washed with complete RPMI 1640 (Gibco®, ThermoFisher Scientific, Inc., Waltham, MA, USA) containing 10% FBS and 100 U/mL penicillin–streptomycin. Red blood cells were lysed with Ammonium-Chloride-Potassium (AKC) lysing buffer (Gibco®, ThermoFisher Scientific, Inc., Waltham, MA, USA). Splenocytes were counted with a hemocytometer under the microscope for cell concentration determination.
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!