Surface staining and intranuclear transcription factor staining were performed as previously described44 (link)45 (link). Briefly, isolated mononuclear cells were incubated with different cocktails of mAbs for surface antigens in 100 μL PBS containing 3% (v/v) FBS at room temperature for 30 minutes. Intranuclear transcription factor staining was performed according to the Foxp3 staining protocol (00-5523-00, eBioscience) after staining with the indicated surface mAbs. Cells were treated with Fixation/Permeabilization Buffer for 1 hour, and subsequently incubated with anti-BCL-6 mAb for 40 minutes at room temperature. Immunostained cells were collected and analysed on a BD FACS Canto II flow cytometer (BD Biosciences, USA). Data were processed using the FlowJo 7.6.1 software.
Foxp3 staining protocol
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Foxp3 staining protocol is a laboratory procedure used to detect and quantify Foxp3-expressing cells, which are a subset of T cells that play a crucial role in regulating the immune system. The protocol involves the use of specific antibodies and reagents to label and identify these cells within a sample, typically obtained from blood, tissue, or other biological sources. The core function of this protocol is to provide researchers and clinicians with a standardized method for analyzing the presence and distribution of Foxp3-positive cells, which is valuable for understanding various immune-related processes and disorders.
Automatically generated - may contain errors
Lab products found in correlation
Facscanto 2 flow cytometer, by BD (1 mentions)
Filipin 3, by Cayman Chemical (1 mentions)
Bodipy, by Thermo Fisher Scientific (1 mentions)
Phosflow lyse fix buffer, by BD (1 mentions)
Phosflow perm buffer 3, by BD (1 mentions)
Facscanto 2, by BD (1 mentions)
Cytoflex s, by Beckman Coulter (1 mentions)
2 protocols using foxp3 staining protocol
1
Multiparameter Flow Cytometry Staining
2
Comprehensive Immune Cell Profiling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Spleens and lymph nodes were mashed through a 70 μm cell strainer after isolation. For the staining of surface markers, cells were stained at 4°C for 30 min in staining buffer [PBS with 2% (vol/vol) foetal bovine serum (FBS)] in which we diluted the antibodies. Intracellular transcription factors were stained for by following the FoxP3 staining protocol (eBioscience, Santa Clara, CA, US.). Filipin III (Cayman Chemicals, Ann Arbor, MI, USA) and Bodipy™ (Thermo Fisher, Waltham, MA, USA) staining for cellular lipids (details in Supplementary material online , Experimental Procedures) was quantified in CD4+CD25+ Treg cells, of which ±95% expressed FoxP3 in SLOs (data not shown). For phosphorylated proteins, cells were fixed with BD Phosflow™ Lyse/Fix Buffer (BD Biosciences) and subsequently permeabilized with Phosflow Perm buffer III (BD Biosciences, Franklin Lakes, NJ, USA). Flow cytometric analysis was performed on a FACSCantoII (BD Biosciences) or a Cytoflex S (Beckman Coulter, Brea, CA, USA) and data were analysed using Flowjo software (TreeStar, Ashland, OR, USA).
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!