Immunofluorescent analysis of NFAT3 using anti-NFAT3 polyclonal antibody (Pierce Product# PA1-021) (shown in green) in HeLa cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were then blocked with 1% Blocker BSA (Pierce Product #37525) for 15 minutes at room temperature. Cells were probed with a rabbit polyclonal antibody recognizing NFAT3 (Pierce Product# PA1-021), at a dilution of 1:100 for at least 1 hour at room temperature. Cells were washed with PBS and incubated with DyLight 488 goat-anti-rabbit secondary antibody (Pierce Product# 35552) at a dilution of 1:400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye (Pierce Product# 62249). Images were taken on a Thermo Scientific ArrayScan at 20X magnification.
|Tested species reactivity||Human|
|Published species reactivity||Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues C R(887) D L S G F P A P P G E E P P A(902) of human NFAT3.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunoprecipitation (IP)||3 µg|
|Western Blot (WB)||1:1000|
* 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 1 publications below|
PA1-021 detects nuclear factor of activated T-cells 3 (NFAT3) from human cells.
PA1-021 has been successfully used in Western blot immunohistochemistry, immunofluorescence, and immunoprecipitation procedures. By Western blot, this antibody detects an ~150 kDa protein representing NFAT3 in HEK293 cells transfected with the human NFAT3 gene. Immunohistochemical staining of NFAT3 in HEK293 cells transfected with the human NFAT3 gene results in cytoplasmic staining.
The PA1-021 immunogen is a synthetic peptide corresponding to residues C R(887) D L S G F P A P P G E E P P A(902) of human NFAT3. This sequence is 94% conserved with mouse NFAT3. The PA1-021 immunizing peptide (Cat. # PEP-122) is available for use in neutralization and control experiments.
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.
Phosphorylation of NFATc4 by p38 mitogen-activated protein kinases.
PA1-021 was used in western blot to study the function of nuclear factor of activated T cells (NFAT) in PPARgamma gene expression and adipocyte differentiation.
|Yang TT,Xiong Q,Enslen H,Davis RJ,Chow CW||Molecular and cellular biology (22:3892)||2002|