44-944G

antibody from Invitrogen Antibodies

Targeting: NFATC2 NF-ATP, NFAT1, NFATp

Provider product page for 44-944G

Western blot

Immunocytochemistry

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [5]
Comments [0]
Validations
Western blot [2]
Immunocytochemistry [1]
Flow cytometry [1]
Other assay [1]

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Product number: 44-944G - Provider product page
Provider: Invitrogen Antibodies
Product name: Phospho-NFATC2 (Ser54) Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Reactivity: Human, Mouse
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Storage: -20°C

NFATC2 protein structure - 44-944G shown in red.

Submitted references Evidence for reprogramming of monocytes into reparative alveolar macrophages in vivo by targeting PDE4b.

Rochford I, Joshi JC, Rayees S, Anwar M, Akhter MZ, Yalagala L, Banerjee S, Mehta D
American journal of physiology. Lung cellular and molecular physiology 2021 Oct 1;321(4):L686-L702

Overexpression of DSCR1 prevents proliferation and predicts favorable prognosis in colorectal cancer patients.

Li WX, Zheng JJ, Zhao G, Lyu CT, Lu WQ
World journal of surgical oncology 2021 Apr 7;19(1):100

STIM1 controls calcineurin/Akt/mTOR/NFATC2-mediated osteoclastogenesis induced by RANKL/M-CSF.

Huang Y, Li Q, Feng Z, Zheng L
Experimental and therapeutic medicine 2020 Aug;20(2):736-747

Regulation of phagocytosis and cytokine secretion by store-operated calcium entry in primary isolated murine microglia.

Heo DK, Lim HM, Nam JH, Lee MG, Kim JY
Cellular signalling 2015 Jan;27(1):177-86

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 2008 Nov 15;112(10):4109-16

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Phospho-NFATC2 (Ser54) using a polyclonal antibody (Product # 44-944G).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of NIH/3T3 (Lane 1), NIH/3T3 treated for 20 minutes with 200 nM of PMA (Lane 2), Jurkat (Lane 3) and Jurkat treated for 20 minutes with 200 nM of PMA (Lane 4). The blots were probed with Anti-NFAT1 (pS54) Rabbit Polyclonal Antibody (Product # 44-944G, 1:500-1:2000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate (Product # G-21234, 1:5000 dilution). A 70 kDa band corresponding to NFAT1 (pS54) was enhanced upon the treatment across the cell lines tested. An additional band was observed at 130 kDa. Known quantity of protein samples were electrophoresed using Novex® NuPAGE®10 % Bis-Tris gel (Product # NP0301BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with PierceTM Power Blotter System (Product # 22834) The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of NFAT1 [pSer54] was done on HeLa cells treated with PMA (200nM, 20 minutes). Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with NFAT1 [pSer54] Rabbit Polyclonal Antibody (44944G, red histogram) or with rabbit isotype control (pink histogram) at 3-5 ug/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Goat Anti-Rabbit Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 3 Overexpression of DSCR1-4 showed tumor suppressive function in CRC cells in vitro. a Western blot showed that overexpression of DSCR1-4 increased p21, p16, and hyperphosphorylation of NFAT1 and NFAT2, while decreased cyclin D1, CDK2, and CDK4. b CCK8 assays showed that overexpression of DSCR1-4 inhibited cell proliferation of CRC cells. Data are mean +- SEM of 5 replicates. c Overexpression of DSCR1-4 inhibited cell colony formation of CRC cells. Data are mean +- SEM of 3 replicates. d Overexpression of DSCR1-4 induced senescence of CRC cells by SA-beta-gal staining. Two-tailed Student t test, * p < 0.05, ** p < 0.01, *** p < 0.001