Naïve CD4+ T cells from C57BL/6 mice were purified by naïve CD4+ T cell negative selection kit (Miltenyi Biotec), and were activated by plate-bound anti-CD3 and anti-CD28 (2 μg ml−1 each) under either neutral condition (Th0) or Th1 differentiating condition in the presence of IL-27 (Th1+IL-27) for 2 days. Cells were rested for additional 3 days in the presence of 10 ng ml−1 of IL-2 and were restimulated with 0.1 μg ml−1 of plate-bound anti-CD3 and anti-CD28 for 24 hours before they were subjected to chromatin preparation for the ChIP analysis. Chromatin fraction preparation and chromatin IP were performed using SimpleChIP™ Enzymatic Chromatin IP Kit (Cell Signaling Technology). Antibody against NFIL3 (C-18) was purchased from Santa Cruz Biotechnology; anti-acetylated Histone 3 antibody (Cat# 06-599) was purchased from Millipore; and an anti-trimethylated Histone Lysine 4 antibody (Cat# ab8580) was purchased from Abcam.
Na ve cd4 t cell negative selection kit
Manufactured by Miltenyi Biotec
The Naïve CD4+ T cell negative selection kit is a laboratory equipment product designed for the isolation of naïve CD4+ T cells from human peripheral blood mononuclear cells. The kit utilizes a magnetic bead-based negative selection method to enrich the target cell population while removing unwanted cell types.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using na ve cd4 t cell negative selection kit
1
Naïve CD4+ T Cell Differentiation
2
Naïve CD4+ T Cell Differentiation
3
CD4+ T Cell Immune Synapse Imaging
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Primary human T cells were isolated from peripheral blood using Lymphoprep (Stemcell) followed by a naïve CD4 T cell negative selection kit (Miltenyi). 1X10 7 naïve human CD4 T cells were transfected with 2 µg CD3ζ-mEOS3 using an Amaxa system (Lonza). Immune synapses were formed against activating coverslips coated with anti-CD3 (2 µg/mL) and anti-CD28 (5 µg/mL). Immune synapses were allowed to form for 5 minutes and were then pH-shift fixed (3% paraformaldehyde (PFA)-kPIPES at 80 mM, pH 6.8 for 5 minutes followed by 3%-PFA Borax at 100 mM for 10 minutes). Cells were imaged in phosphate buffered saline (PBS).
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!