Immunofluorescence analysis of Phospho-NFAT1 (Ser54) was done on 70% confluent log phase HeLa cells treated with 200nM of PMA for 20 minutes. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with phospho-NFAT1 (Ser54) Rabbit Polyclonal Antibody (Product # 44-944G) at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing cytoplasmic and nuclear localization. Panel e is untreated cells with no signal. Panel f is no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from a region of mouse NFAT1 that contains serine 54.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 50% glycerol, 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:20|
|Western Blot (WB)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 2 publications below|
The nuclear factor of activated T-cells (NFAT) transcription complex is required for the expression of a group of proteins that collectively regulate the immune response. Four NFAT proteins, encoded on separate genes and expressed as several splice variants, have been described: NFAT1 (also known as NFATp or NFATc2), NFAT2 (NFATc or NFATc1), NFAT3, and NFAT4 (NFATx or NFATc3). These proteins show a low level of sequence similarity with the Dorsal/Rel/NFkB family of transcription factors. Another NFAT-related protein termed NFAT5 differs from isoforms 1-4 in that it lacks many of the Fos/Jun contact sites observed in its predecessors and its subcellular localization is not calcineurin-dependent.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Regulation of phagocytosis and cytokine secretion by store-operated calcium entry in primary isolated murine microglia.
44-944G was used in western blot to determine that SOCE regulates UDP-induced phagocytosis and LPS-stimulated cytokine secretion in microglia
|Heo DK,Lim HM,Nam JH,Lee MG,Kim JY||Cellular signalling (27:177)||2015|
|Mouse||Not Cited||NK cell-activating receptors require PKC-theta for sustained signaling, transcriptional activation, and IFN-gamma secretion.||Tassi I,Cella M,Presti R,Colucci A,Gilfillan S,Littman DR,Colonna M||Blood (112:4109)||2008|