NFAT1 Antibody | Affinity Biosciences

WB
Cites

1/7

Western blot analysis of extracts from Hela, using NFATC2 Antibody.

Western blot analysis of extracts from A549 cells(heat shock treatment), using NFAT1 Antibody.

Fig.

Figure 5.

Figure 9.

FIGURE 5 Aβ oligomer-induced expression levels of BACE1 and inactive NFAT1 by interference with Nav1.

Product:	NFAT1 Antibody
Catalog:	DF7189
Description:	Rabbit polyclonal antibody to NFAT1
Application:	WB
Reactivity:	Human, Mouse, Rat
Prediction:	Pig, Bovine, Horse, Rabbit, Dog, Chicken, Xenopus
Mol.Wt.:	100-150kD; 100kD(Calculated).
Uniprot:	Q13469
RRID:	AB_2839141

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Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific.

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MSDS

Data Sheet

COA

Protocols

WB Handbook

IHC Protocol

IF/ICC Protocol

Product Info

Source:

Rabbit

Application:

WB 1:500-1:1000
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:

Human,Mouse,Rat

Prediction:

Pig(100%), Bovine(100%), Horse(100%), Rabbit(100%), Dog(100%), Chicken(100%), Xenopus(88%)

Clonality:

Polyclonal

Specificity:

NFAT1 Antibody detects endogenous levels of total NFAT1.

RRID:

AB_2839141
Cite Format: Affinity Biosciences Cat# DF7189, RRID:AB_2839141.

Conjugate:

Unconjugated.

Purification:

The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).

Storage:

Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.

Alias:

Fold/Unfold

AI607462; cytoplasmic 2; KIAA0611; NF ATc2; NF ATp; NF-ATc2; NF-ATp; NFAC2_HUMAN; NFAT 1; NFAT pre existing subunit; NFAT pre-existing subunit; NFAT transcription complex, preexisting component; NFAT1; NFAT1-D; NFATc2; NFATp; Nuclear factor of activated T cells cytoplasmic 2; Nuclear factor of activated T cells cytoplasmic calcineurin dependent 2; Nuclear factor of activated T cells pre-existing component; Nuclear factor of activated T cells, preexisting component; Nuclear factor of activated T-cells; Preexisting nuclear factor of activated T cells 2; T cell transcription factor NFAT 1; T-cell transcription factor NFAT1;

Immunogens

Immunogen:

Uniprot:

Q13469(NFAC2_HUMAN) >>Visit HPA database.

Gene(ID):

NFATC2(4773)

Expression:

Q13469 NFAC2_HUMAN:

Expressed in thymus, spleen, heart, testis, brain, placenta, muscle and pancreas. Isoform 1 is highly expressed in the small intestine, heart, testis, prostate, thymus, placenta and thyroid. Isoform 3 is highly expressed in stomach, uterus, placenta, trachea and thyroid.

Description:

This gene is a member of the nuclear factor of activated T cells (NFAT) family. The product of this gene is a DNA-binding protein with a REL-homology region (RHR) and an NFAT-homology region (NHR). This protein is present in the cytosol and only translocates to the nucleus upon T cell receptor (TCR) stimulation, where it becomes a member of the nuclear factors of activated T cells transcription complex. This complex plays a central role in inducing gene transcription during the immune response. Alternate transcriptional splice variants encoding different isoforms have been characterized.

Sequence:

Q13469 NFAC2_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MNAPERQPQPDGGDAPGHEPGGSPQDELDFSILFDYEYLNPNEEEPNAHKVASPPSGPAYPDDVLDYGLKPYSPLASLSGEPPGRFGEPDRVGPQKFLSAAKPAGASGLSPRIEITPSHELIQAVGPLRMRDAGLLVEQPPLAGVAASPRFTLPVPGFEGYREPLCLSPASSGSSASFISDTFSPYTSPCVSPNNGGPDDLCPQFQNIPAHYSPRTSPIMSPRTSLAEDSCLGRHSPVPRPASRSSSPGAKRRHSCAEALVALPPGASPQRSRSPSPQPSSHVAPQDHGSPAGYPPVAGSAVIMDALNSLATDSPCGIPPKMWKTSPDPSPVSAAPSKAGLPRHIYPAVEFLGPCEQGERRNSAPESILLVPPTWPKPLVPAIPICSIPVTASLPPLEWPLSSQSGSYELRIEVQPKPHHRAHYETEGSRGAVKAPTGGHPVVQLHGYMENKPLGLQIFIGTADERILKPHAFYQVHRITGKTVTTTSYEKIVGNTKVLEIPLEPKNNMRATIDCAGILKLRNADIELRKGETDIGRKNTRVRLVFRVHIPESSGRIVSLQTASNPIECSQRSAHELPMVERQDTDSCLVYGGQQMILTGQNFTSESKVVFTEKTTDGQQIWEMEATVDKDKSQPNMLFVEIPEYRNKHIRTPVKVNFYVINGKRKRSQPQHFTYHPVPAIKTEPTDEYDPTLICSPTHGGLGSQPYYPQHPMVAESPSCLVATMAPCQQFRTGLSSPDARYQQQNPAAVLYQRSKSLSPSLLGYQQPALMAAPLSLADAHRSVLVHAGSQGQSSALLHPSPTNQQASPVIHYSPTNQQLRCGSHQEFQHIMYCENFAPGTTRPGPPPVSQGQRLSPGSYPTVIQQQNATSQRAAKNGPPVSDQKEVLPAGVTIKQEQNLDQTYLDDVNEIIRKEFSGPPARNQT

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species

Results

Score

Pig

100

Horse

100

Bovine

100

Dog

100

Chicken

100

Rabbit

100

Xenopus

Sheep

Zebrafish

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q13469 As Substrate

Q13469

Site	PTM Type	Enzyme	Source
S23	Phosphorylation		Uniprot
S31	Phosphorylation	Q86Y07 (VRK2)	Uniprot
Y36	Phosphorylation		Uniprot
Y38	Phosphorylation		Uniprot
S53	Phosphorylation	Q05513 (PRKCZ)	Uniprot
S56	Phosphorylation	Q05513 (PRKCZ)	Uniprot
K70	Ubiquitination		Uniprot
S73	Phosphorylation		Uniprot
S79	Phosphorylation		Uniprot
K96	Ubiquitination		Uniprot
S99	Phosphorylation		Uniprot
K102	Ubiquitination		Uniprot
S107	Phosphorylation		Uniprot
S110	Phosphorylation		Uniprot
T116	Phosphorylation		Uniprot
S148	Phosphorylation		Uniprot
R150	Methylation		Uniprot
S171	Phosphorylation		Uniprot
S172	Phosphorylation		Uniprot
S174	Phosphorylation		Uniprot
S175	Phosphorylation		Uniprot
S177	Phosphorylation		Uniprot
Y212	Phosphorylation		Uniprot
S213	Phosphorylation		Uniprot
S217	Phosphorylation		Uniprot
S221	Phosphorylation		Uniprot
S225	Phosphorylation		Uniprot
S230	Phosphorylation		Uniprot
S236	Phosphorylation		Uniprot
S243	Phosphorylation		Uniprot
S245	Phosphorylation		Uniprot
S247	Phosphorylation		Uniprot
S255	Phosphorylation		Uniprot
S268	Phosphorylation		Uniprot
S274	Phosphorylation		Uniprot
S276	Phosphorylation		Uniprot
S280	Phosphorylation		Uniprot
T325	Phosphorylation		Uniprot
S326	Phosphorylation		Uniprot
S330	Phosphorylation		Uniprot
S333	Phosphorylation		Uniprot
S337	Phosphorylation		Uniprot
K338	Ubiquitination		Uniprot
Y346	Phosphorylation		Uniprot
S363	Phosphorylation		Uniprot
S367	Phosphorylation		Uniprot
T374	Phosphorylation		Uniprot
K482	Ubiquitination		Uniprot
T483	Phosphorylation		Uniprot
T485	Phosphorylation		Uniprot
K491	Ubiquitination		Uniprot
K497	Ubiquitination		Uniprot
K520	Ubiquitination		Uniprot
K530	Ubiquitination		Uniprot
T585	Phosphorylation		Uniprot
T599	Phosphorylation		Uniprot
S605	Phosphorylation		Uniprot
T652	Phosphorylation		Uniprot
S736	Phosphorylation		Uniprot
S737	Phosphorylation		Uniprot
Y752	Phosphorylation		Uniprot
S755	Phosphorylation		Uniprot
S757	Phosphorylation		Uniprot
S759	Phosphorylation		Uniprot
S761	Phosphorylation		Uniprot
S776	Phosphorylation		Uniprot
S790	Phosphorylation		Uniprot
S801	Phosphorylation		Uniprot
S808	Phosphorylation		Uniprot
S814	Phosphorylation		Uniprot
S856	Phosphorylation		Uniprot
S859	Phosphorylation		Uniprot
Y860	Phosphorylation		Uniprot
S882	Phosphorylation		Uniprot
K885	Ubiquitination		Uniprot
T893	Phosphorylation		Uniprot
K895	Sumoylation		Uniprot
Y904	Phosphorylation		Uniprot

Research Backgrounds

Q13469_NFAC2_HUMAN

Function:

Plays a role in the inducible expression of cytokine genes in T-cells, especially in the induction of the IL-2, IL-3, IL-4, TNF-alpha or GM-CSF. Promotes invasive migration through the activation of GPC6 expression and WNT5A signaling pathway.

PTMs:

In resting cells, phosphorylated by NFATC-kinase on at least 18 sites in the 99-363 region. Upon cell stimulation, all these sites except Ser-243 are dephosphorylated by calcineurin. Dephosphorylation induces a conformational change that simultaneously exposes an NLS and masks an NES, which results in nuclear localization. Simultaneously, Ser-53 or Ser-56 is phosphorylated; which is required for full transcriptional activity.

Ubiquitinated in endothelial cells by RNF213 downstream of the non-canonical Wnt signaling pathway, leading to its degradation by the proteasome.

Subcellular Location:

Cytoplasm. Nucleus.
Note: Cytoplasmic for the phosphorylated form and nuclear after activation that is controlled by calcineurin-mediated dephosphorylation. Rapid nuclear exit of NFATC is thought to be one mechanism by which cells distinguish between sustained and transient calcium signals. The subcellular localization of NFATC plays a key role in the regulation of gene transcription.

Tissue Specificity:

Subunit Structure:

Member of the multicomponent NFATC transcription complex that consists of at least two components, a pre-existing cytoplasmic component NFATC2 and an inducible nuclear component NFATC1. Other members such as NFATC4, NFATC3 or members of the activating protein-1 family, MAF, GATA4 and Cbp/p300 can also bind the complex. The phosphorylated form specifically interacts with XPO1; which mediates nuclear export. NFATC proteins bind to DNA as monomers. Interacts with NFATC2IP (By similarity). Interacts with FOXP3. Interacts with TBX21 ('Thr-303' phosphorylated form) (By similarity). Interacts with KAT2A (By similarity). Interacts with HOMER2 and HOMER3; this interaction competes with calcineurin/PPP3CA-binding and hence prevents NFATC2 dephosphorylation and activation. Interacts with protein phosphatase PPP3CA/calcineurin A.

Family&Domains:

the 9aaTAD motif is a transactivation domain present in a large number of yeast and animal transcription factors.

Rel Similarity Domain (RSD) allows DNA-binding and cooperative interactions with AP1 factors.

Research Fields

· Cellular Processes > Cell growth and death > Cellular senescence. (View pathway)

· Environmental Information Processing > Signal transduction > cGMP-PKG signaling pathway. (View pathway)

· Environmental Information Processing > Signal transduction > Wnt signaling pathway. (View pathway)

· Human Diseases > Infectious diseases: Viral > Hepatitis B.

· Human Diseases > Infectious diseases: Viral > HTLV-I infection.

· Organismal Systems > Development > Axon guidance. (View pathway)

· Organismal Systems > Development > Osteoclast differentiation. (View pathway)

· Organismal Systems > Immune system > Natural killer cell mediated cytotoxicity. (View pathway)

· Organismal Systems > Immune system > Th1 and Th2 cell differentiation. (View pathway)

· Organismal Systems > Immune system > Th17 cell differentiation. (View pathway)

· Organismal Systems > Immune system > T cell receptor signaling pathway. (View pathway)

· Organismal Systems > Immune system > B cell receptor signaling pathway. (View pathway)

· Organismal Systems > Endocrine system > Oxytocin signaling pathway.

References

1). Reducing Nav1.6 expression attenuates the pathogenesis of Alzheimer's disease by suppressing BACE1 transcription. Aging Cell, 2022 [IF=7.8]

Application: WB Species: Mouse Sample:

FIGURE 5 Aβ oligomer-induced expression levels of BACE1 and inactive NFAT1 by interference with Nav1.6, TTX, EGTA, and KB-R7943. Representative immunoblots (a) and densitometry analysis of P-NFAT1 (inactive) (b), NFAT1 (active) (c), BACE1 (d), and Nav1.6 (e) expression levels in SH-SY5Y cells after treatment with siNav1.6, 1 μM TTX, 0.5 mM EGTA, and 5 μM KB-R7943 under induced Aβ oligomers condition. (f) Relative mRNA expression of RT-qPCR showing the expression level of BACE1 in the SH-SY5Y cells after treatment with siNav1.6, 1 μM TTX, 0.5 mM EGTA, and 5 μM KB-R7943 under induced Aβ oligomers condition. (g) Intracellular calcium levels in SH-SY5Y cell after treatment with NC, siNav1.6, 1 μM TTX, 0.5 mM EGTA, and 5 μM KB-R7943 under induced Aβ oligomers condition. (h) SH-SY5Y cells were either treated with 1 μM TTX, 0.5 mM EGTA, and 5 μM KB-R7943 or transfected with Nav1.6 siRNA in the presence of Aβ oligomers and immunostained for NFAT1 and DAPI. Scale bar: 100 μm. (i) The ratio of nuclear NFAT1 to the cytoplasmic NFAT1. Here, P-NFAT1, BACE1, and Nav1.6 were normalized to γ-tubulin in the Western blot, whereas BACE1 was normalized to ACTIN in the RT-PCR. Data are presented as mean ± SEM. *p < 0.05, **p < 0.01, **p < 0.001. Representative immunoblots (j) and densitometry analysis (k) of Nav1.6, P-NFAT1, and NFAT1 protein expression in the brain of mice (WT and APP/PS1 treated with siNav1.6 or NC), n = 5 mice/group. (l) The proposed flowchart cycle depicting interference of Nav1.6’s effect on the progression of AD. Interference of Nav1.6 can reduce the Aβ oligomers-dependent transcription of BACE1, which then relieves intracellular calcium overload by inhibiting sodium-calcium reverse exchange channel and leads to an increase in non-activated NFAT1 expression levels. The reduced transcription of BACE1, in turn, decreases Aβ production and slows the progression of AD

Application: IF/ICC Species: Mouse Sample:

2). Dehydroepiandrosterone attenuates pulmonary artery and right ventricular remodeling in a rat model of pulmonary hypertension due to left heart failure. LIFE SCIENCES, 2019 (PubMed: 30605649) [IF=6.1]

Application: WB Species: rat Sample: lung

Fig. 6. |DHEA suppressed STAT3/NFAT signal pathway in lung. A, PY750-STAT3 and STAT3; B, NFATc2; C, Pim-1; D, Survivin. The top shows representative immunoblots, and the bottom shows the densitometric assessment. The values are means ± SE, n = 5 rats per group; *P < 0.05 versus the sham group, #P < 0.05 versus the PH-LHF group.

3). Effects and Mechanisms of Rhus chinensis Mill. Fruits on Suppressing RANKL-Induced Osteoclastogenesis by Network Pharmacology and Validation in RAW264.7 Cells. Nutrients, 2022 (PubMed: 35267996) [IF=5.9]

Application: WB Species: Sample: RAW264.7 cells

Figure 9.| Effects of the ethanolic extract from the R. chinensis fruits on RANKL-induced osteoclastogenesis on c-Fos and NFATc1 proteins in RAW264.7 cells. (a) Western blot analysis of c-Fos and proteins

4). Wnt5a regulates Ameloblastoma Cell Migration by modulating Mitochondrial and Cytoskeletal Dynamics. Journal of Cancer, 2020 (PubMed: 32742496) [IF=3.9]

Application: WB Species: human Sample: AM-1 cells

Figure 5. |Wnt5a up-regulates mitochondria-cytoskeleton co-localization through mediating Coro1A expression. (A) Western blot detected nuclear transfactor NFATc2 and Coro1A protein expression. Bar charts demonstrating quantitative significance results from 3 to 5 independent experiments. (B) Western blot demonstrating Coro1A protein expression changes of AM-1 cells treated with only human recombinant Wnt5a compared with cells in the presence of both rhWnt5a and FK506 (200 nm).

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NFAT1 Antibody - #DF7189