FC-FP: FC analysis of fixed/permeabilized human peripheral blood mononuclear cells, untreated (left column) or treated with Rapid-Act T Cell Activation Kit (Human, Anti-CD3/CD28) #88179 (15 min; right column), using #38950 or concentration-matched Rabbit (DA1E) Monoclonal Antibody IgG Isotype Control #3900 (bottom row), co-stained with CD3 (UCHT1) Mouse Monoclonal Antibody (PE Conjugate) #46233. Anti-rabbit IgG (H+L), F(ab’) Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody

Mechanisms of regulatory activity of the NFAT transcription factor family and calcium signaling.

The NFAT family (NFAT1–4) are transcription factors possessing a Rel homologous domain, and they work in conjunction with AP-1 to regulate the transcription of immune response-related genes. Calcium signaling is central to their activity regulation. NFAT1, highly phosphorylated in the quiescent phase, is dephosphorylated by activated calcineurin in response to increased intracellular calcium concentration and translocates to the nucleus. Conversely, rephosphorylation by GSK-3 and CK1 in response to decreased calcium concentration terminates the signal. Immunosuppressants such as cyclosporine A and FK506 are known to inhibit calcineurin activity, thereby preventing NFAT nuclear translocation. This pathway is a crucial molecular basis for T cell activation. #38950 is suitable for WB, IP, and FC-FP using human samples.

IF-F: Confocal IF analysis of fixed frozen mouse lung labeled with #61741 (green). Free secondary binding sites were then blocked with Rabbit (DA1E) Monoclonal Antibody IgG Isotype Control #3900 prior to co-labeling with CD45 (D3F8Q) Rabbit Monoclonal Antibody (Alexa Fluor® 555 Conjugate) #19581 (gray pseudocolor), CD31 (PECAM-1) (F2N3M) Rabbit Monoclonal Antibody (Alexa Fluor® 647 Conjugate) #34624 (red), and ProLong Gold Antifade Reagent with DAPI #8961 (blue).

Mechanisms of regulatory activity of the NFAT transcription
factor family and calcium signaling. 

Krüppel-like factors (KLFs) are a group of zinc finger-type transcriptional regulators homologous to Krüppel, a somite formation factor in Drosophila. Currently, 17 types have been identified in mammals, regulating a wide range of life phenomena including development, differentiation, proliferation, and stress response. Dysfunction of these KLF family members is involved in the pathogenesis of various diseases, such as metabolic disorders and cancer. KLF2, in particular, is a key factor in the differentiation and maintenance of function of immune cells and vascular endothelial cells. KLF2 exerts anti-inflammatory effects by suppressing the expression of inflammatory genes via the NF-κB pathway, playing a crucial role in regulating immune responses and maintaining tissue homeostasis. #61741 is suitable for WB, IHC-P, and IF-F using mouse samples.

• Get precise results with Q5’s high fidelity (~280x Taq)
• Reduce turnaround time through fast cycling conditions, with extension times of 5-15 seconds/kb
• Optimized formulation enables robust, reliable amplification of diverse targets (from high AT or high GC) up to 20 kb
• Improve reaction specificity with a hot start aptamer that allows a convenient room temperature setup
• Compatible with a 62°C universal annealing temperature protocol. Learn more in our application note
• Also available: Q5 standalone polymerase (NEB #M0491, NEB #M0493, NEB #M0515) and master mix formats (NEB #M0492, NEB #M0494, NEB #M0578, NEB #M0580)