NFAT1 Recombinant Rabbit Monoclonal Antibody [JA11-08]
Overview
Product Name
NFAT1 Recombinant Rabbit Monoclonal Antibody [JA11-08]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human NFAT1 aa 21-70 / 925.
Species Reactivity
Human
Validated Applications
WB, IF-Tissue, IHC-P
Molecular Weight
Predicted band size: 100 kDa
Positive Control
Jurkat cell lysate, Ramos cell lysate, Raji cell lysate, Daudi cell lysate, human spleen tissue, human NK-T-cell lymphoma tissue, human colon cancer tissue.
Conjugation
unconjugated
Clone Number
JA11-08
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
-
WB
-
1:1,000
-
IF-Tissue
-
1:50-1:200
-
IHC-P
-
1:1,000
Target
Function
The NFAT (nuclear factor of activated T cells) family of transcription factors regulates cytokine expression in T cells. Members of the family include NFATc1 (NFATc), NFATc2 (NFATp), NFATn, NFATc3 (NFAT4, NFATx) and NFATc4 (NFAT3). Recognition of antigen by the T cell receptor (TCR) eventually activates the calcium-dependent protein phosphatase calcineurin. Once activated, calcineurin stimulates the translocation of NFATc1 (cytoplasmic) from the cytoplasm to the nucleus where it associates with NFATn (nuclear). Like NFATc1, NFATc2 resides in the cytoplasm and translocates to the nucleus subsequent to activation of calcineurin. Once in the nucleus, NFATc2 synergizes with AP-1 transcription factors to initiate transcription of cytokine genes. NFATc3 and NFATc4 share 65% sequence identity with other members of the NFAT family. They are similar to NFATc2 in that they also synergize with the AP-1 family of proteins.
Background References
1. Makowski SL et al. A protease-independent function for SPPL3 in NFAT activation. Mol Cell Biol 35:451-67 (2015).
2. Chin-Smith EC et al. Nuclear factor of activated T-cell isoform expression and regulation in human myometrium. Reprod Biol Endocrinol 13:83 (2015).
Tissue Specificity
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.
Post-translational Modification
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.
UNIPROT
Synonyms
AI607462 antibody
cytoplasmic 2 antibody
KIAA0611 antibody
NF ATc2 antibody
NF ATp antibody
NF-ATc2 antibody
NF-ATp antibody
NFAC2_HUMAN antibody
NFAT 1 antibody
NFAT pre existing subunit antibody
ExpandImages
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☑ Relative expression (RE)
Western blot analysis of NFAT1 on different lysates with Rabbit anti-NFAT1 antibody (ET1704-14) at 1/1,000 dilution.
Lane 1: Jurkat cell lysate
Lane 2: Ramos cell lysate
Lane 3: Raji cell lysate
Lane 4: Daudi cell lysate
Lane 5: HeLa cell lysate (negative)
Lysates/proteins at 30 µg/Lane.
Predicted band size: 100 kDa
Observed band size: 120/140 kDa
Exposure time: 10 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1704-14) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-NFAT1 antibody (ET1704-14) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-14) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human NK-T-cell lymphoma tissue with Rabbit anti-NFAT1 antibody (ET1704-14) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-14) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-NFAT1 antibody (ET1704-14) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-14) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Citation
-
Osteopontin isoform c promotes the survival of cisplatin-treated NSCLC cells involving NFATc2-mediated suppression on calcium-induced ROS levels
Journal: BMC Cancer
DOI:
IF: 4.431
Application: IF-cell
Reactivity: Human
Publish date: 2021 Jun
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