Rabbit Anti-phospho-NFATc2 (Ser330) antibody

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NFATC2(phospho Ser330); NFATC2(phospho S330); AI607462; KIAA0611; NF ATp; NF-ATc2; NFAT 1; NFAT pre existing subunit; NFAT transcription complex, preexisting component; NFAT1; NFAT1-D; NFATc2; NFATp; Nuclear factor of activated T cells cytoplasmic 2; Nucl

Cat:

SL5516R

Species Reactivity：

Human,Mouse,Rat,(predicted: Dog,Pig,Cow,Horse,)

Immunogen：

KLH conjugated Synthesised phosphopeptide derived from human NFATc2 around the phosphorylation site of Ser330:DP(p-S)PV

Format：

Liquid

Storage instructions：

Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Concentration：

1mg/ml

Clonality：

Polyclonal

Isotype：

IgG

Applications：

WB=1:500-2000ELISA=1:5000-10000IHC-P=1:100-500Flow-Cyt=1ug/Test（Paraffin sections need to do antigen repair）not yet tested in other applications.optimal dilutions/concentrations should be determined by the end user.

Host：

Rabbit

Product Overview：

Sample: Lane 1: Mouse Thymus tissue lysates Lane 2: Mouse Spleen tissue lysates Lane 3: Human Raji cell lysates Lane 4: Human Jurkat cell lysates Lane 5: Human U251 cell lysates Primary: Anti-phospho-NFATc2 (Ser330) (SL5516R) at 1/1000 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 100 kDaObserved band size: 140 kDa Paraformaldehyde-fixed, paraffin embedded (Rat brain); Antigen retrieval by microwave in sodium citrate buffer (pH6.0) ; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer (3% BSA) at RT for 30min; Antibody incubation with (phospho-NFATc2(Ser330)) Polyclonal Antibody, Unconjugated (SL5516R) at 1:400 overnight at 4°C, followed by conjugation to the secondary antibody (labeled with HRP)and DAB staining.Blank control（black line）:Molt4. Primary Antibody (green line): Rabbit Anti-phospho-NFATc2 (Ser330) antibody (SL5516R) Dilution:1ug/Test; Secondary Antibody（white blue line）: Goat anti-rabbit IgG-AF488Dilution: 0.5ug/Test. Isotype control（orange line）: Normal Rabbit IgGProtocolThe cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃, The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

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Unit:

Price: $256.00

Product PDFs

Datasheet: Down Datasheet

Other Information
Citations
Protein Attributes
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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. [provided by RefSeq, Apr 2012]

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.

Subunit:
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).

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:
Expressed in thymus, spleen, heart, testis, brain, placenta, muscle and pancreas.

Post-translational modifications:
In resting cells, phosphorylated by NFATSLCkinase on at least 18 sites in the 99-365 region. Upon cell stimulation, all these sites except Ser-245 are dephosphorylated by calcineurin. Dephosphorylation induces a conformational change that simultaneously exposes an NLS and masks an NES, which results in nuclear localization. Simultaneously, one site among Ser-53; Ser-54 and Ser-56 is phosphorylated; which is required for full transcriptional activity.

Similarity:
Contains 1 RHD (Rel-like) domain.

SWISS:
Q13469

Gene ID:
4773

Database links:

Entrez Gene: 4773 Human

Entrez Gene: 3619 Mouse

Entrez Gene: 311658 Rat

Omim: 600490 Human

SwissProt: Q13469 Human

SwissProt: Q60591 Mouse

Unigene: 713650 Human

Unigene: 744148 Human

Unigene: 13362 Mouse

Unigene: 33679 Rat

Picture

Paraformaldehyde-fixed, paraffin embedded (Rat brain); Antigen retrieval by microwave in sodium citrate buffer (pH6.0) ; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer (3% BSA) at RT for 30min; Antibody incubation with (phospho-NFATc2(Ser330)) Polyclonal Antibody, Unconjugated (SL5516R) at 1:400 overnight at 4°C, followed by conjugation to the secondary antibody (labeled with HRP)and DAB staining.

Blank control（black line）:Molt4.
Primary Antibody (green line): Rabbit Anti-phospho-NFATc2 (Ser330) antibody (SL5516R)
Dilution:1ug/Test;
Secondary Antibody（white blue line）: Goat anti-rabbit IgG-AF488
Dilution: 0.5ug/Test.
Isotype control（orange line）: Normal Rabbit IgG
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃, The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

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