Naïve CD4 T cells were purified from splenocytes of WT mice using Miltenyi naïve CD4 isolation kit. For Th1 induction in Figure 1A , naïve CD4 T cells were cultured on 24-well plates coated with 1 μg/ml of αCD3/CD28 (Biolegend) plus IL-12 (0.5 ng/ml) and AS1842856 (or DMSO) for 72h. For iTreg induction in Figure 5 , αCD3/CD28 (1 μg/ml) and recombinant mouse PD-L1-Ig chimera or human IgG1-Ig (10 μg/ml) (Biolegend) were used to coat plates. Naïve CD4 T cells were cultured on coated plates plus TGFβ (4 μg/ml) and AS1842856 (or DMSO) for 72h. For the expansion of human Th1 cells in Figure 4 C –D , PBMCs from treatment-naïve MS patients were plated in flasks for 2–4 hrs to remove adherent cells. The suspension cells were then collected and activated with plate-bound αCD3/CD28 plus IL-12 (0.5 ng/ml) and AS1842856 (0.1 μM) or DMSO for 3 days.
Na ve cd4 isolation kit
Manufactured by Miltenyi Biotec
Sourced in Germany
The Naïve CD4 isolation kit is a product designed to isolate naïve CD4+ T cells from human peripheral blood mononuclear cells (PBMCs). The kit utilizes a magnetic bead-based separation process to negatively select for naïve CD4+ T cells, retaining their native phenotype and functionality.
Automatically generated - may contain errors
Lab products found in correlation
αcd3 cd28, by BioLegend (1 mentions)
Recombinant mouse pd l1 ig chimera or human igg1 ig, by BioLegend (1 mentions)
Cellxvivo mouse th17 cell differentiation kit, by R&D Systems (1 mentions)
Facsaria, by BD (1 mentions)
Lymphoprep, by Axis-Shield (1 mentions)
Pan t cell isolation kit, by Miltenyi Biotec (1 mentions)
4 protocols using na ve cd4 isolation kit
1
Induction of Th1 and iTreg Cells from Murine Naive CD4+ T Cells
2
Isolation and Differentiation of Murine TH17 Cells
3
Naive CD4+ T Cell Isolation and Adoptive Transfer
4
Tofacitinib Modulates T Cell Proliferation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PBMCs were isolated from healthy volunteer whole blood samples using Lymphoprep (Axis Shield, Dundee, United Kingdom) according to the manufacturer’s instructions. Pan T cells were isolated using a Pan T Cell isolation kit (Miltenyi Biotec, Bergisch Gladbach, Germany), stimulated with αCD3/αCD28 for 3 days in the presence of tofacitinib (10–500 nM) or dimethyl sulfoxide and the proliferation was measured using a WST-1 assay. For flow cytometric analyses, pan T cells were stained with CFSE prior to stimulation with αCD3/αCD28 for 5 days in the presence of tofacitinib (10–500 nM) or dimethyl sulfoxide. Human naïve CD4+ T cells were isolated from PMBCs using a naïve CD4+ isolation kit (Miltenyi Biotec). For cytokine and RNA measurements the cells were stimulated with αCD3/αCD28 and IL-12 (Th1), IL-4 (Th2) or IL-6, IL-1β, TGF-β, and IL-23 (Th17) for 5 days. Tofacitinib or dimethyl sulfoxide was either added at the start of the experiment or after the differentiation (day 5) and cells were restimulated with PHA. For flow cytometric analysis naïve CD4+ T cells were stained with CFSE prior to differentiation as mentioned 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!