Log In Sign up
The largest database of trusted experimental protocols

Nfatc4

Manufactured by Santa Cruz Biotechnology
Visit Supplier
Sourced in United States, Germany

NFATc4 is a protein that plays a role in the regulation of gene expression in response to calcium signaling. It is a member of the Nuclear Factor of Activated T-cells (NFAT) family of transcription factors. NFATc4 functions as a key regulator of cellular processes, but a detailed description of its core function cannot be provided while maintaining an unbiased and factual approach without extrapolation.

Automatically generated - may contain errors

Lab products found in correlation

Nfatc2, by Cell Signaling Technology (2 mentions) Nfatc3, by Cell Signaling Technology (2 mentions) Stim1, by BD (1 mentions) Stim2, by Cell Signaling Technology (1 mentions) Trpc1, by Alomone (1 mentions) Anti glyceraldehyde 3 phosphate dehydrogenase gapdh monoclonal antibody, by Santa Cruz Biotechnology (1 mentions) Trpc3, by Alomone (1 mentions) Trpc6, by Alomone (1 mentions) α tubulin, by Merck Group (1 mentions) Nfatc3, by Santa Cruz Biotechnology (1 mentions) C myc, by Santa Cruz Biotechnology (1 mentions) Nfatc1, by Cell Signaling Technology (1 mentions) Gapdh, by Santa Cruz Biotechnology (1 mentions) Bq123, by Merck Group (1 mentions) Angiotensin 2, by Merck Group (1 mentions) Endothelin 1, by Merck Group (1 mentions) Anti nanog, by Abcam (1 mentions) Anti oct3 4, by Santa Cruz Biotechnology (1 mentions) Bq788, by Merck Group (1 mentions) Anti ssea 3, by Merck Group (1 mentions)

8 protocols using nfatc4

1

Cardiac Protein Expression Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
All protein samples were extracted from mice hearts. Protein concentrations were determined using a Bradford assay with bovine serum albumin (BSA) as a standard. To determine the protein expression of STIM1 (BD Biosciences, CA, USA), STIM2 (Cell Signaling Technology, MA, USA), TRPC1, TRPC3, TRPC4, and TRPC6 (Alomone Labs, Jerusalem, Israel), pNFATc4 (rabbit anti-mouse polyclonal antibody raised against short amino acid sequence containing Ser 168 and 170 dually phosphorylated NFATc4 of human origin, Santa Cruz Biotechnology, TX, USA), NFATc4 (rabbit anti-mouse polyclonal antibody raised against amino acids 125–198 of NFATc4 human origin, Santa Cruz Biotechnology, TX, USA), samples (50 μg) were run on 6% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), transferred to polyvinylidene fluoride (PVDF) membranes, and rocked at 4°C for 1 h in blocking buffer (0.1% Tween 20 and 1% BSA in Tris-buffered saline). An enhanced chemiluminescence (ECL)-detection system was used to detect the bound antibodies. An anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) monoclonal antibody (Santa Cruz Biotechnology, TX, USA) was used as an internal loading control.
Ohba T., Watanabe H., Murakami M., Iino K., Adachi T., Baba Y., Kurosaki T., Ono K, & Ito H. (2017). Stromal interaction molecule 1 haploinsufficiency causes maladaptive response to pressure overload. PLoS ONE, 12(11), e0187950.
+ Open protocol
+ Expand
2

Western Blot Analysis of NFAT Proteins

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Western blotting was performed as described previously [67 (link)]. We used the following primary antibodies for our study: NFATc1 (8032S; Cell Signaling Tech), NFATc2 (sc-7296; Santa Cruz Biotech), NFATc3 (sc-8405; Santa Cruz Biotech), NFATc4 (sc-13036; Santa Cruz Biotech), C-MYC (sc-40; Santa Cruz Biotech), α-tubulin (T9026; Sigma), GAPDH (FL-335; Santa Cruz Biotech), and DDK (TA50011-100; OriGene).
Lee S.H., Kieu C., Martin C.E., Han J., Chen W., Kim J.S., Kang M.K., Kim R.H., Park N.H., Kim Y, & Shin K.H. (2019). NFATc3 plays an oncogenic role in oral/oropharyngeal squamous cell carcinomas by promoting cancer stemness via expression of OCT4. Oncotarget, 10(23), 2306-2319.
+ Open protocol
+ Expand
3

Immunofluorescence Staining of Pluripotency and Cardiac Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols
Immunofluorescence staining for pluripotency of iPSCs was performed by using the following primary antibodies: anti‐NANOG (Abcam), anti‐OCT3/4 (Santa Cruz), anti–SSEA 3 (Millipore), anti–SSEA 4 (Millipore), anti–Tra‐1‐60 (Millipore), and anti–Tra‐1‐81 (Millipore). In immunofluorescence staining for cardiac markers, monoclonal anti–α‐actinin (Sigma), monoclonal anti‐cTnT (Thermo Scientific), polyclonal anti‐cTnT (Santa Cruz), monoclonal anti–myosin light chain (MLC)2a (Synaptic Systems), polyclonal anti‐MLC2v (ProteinTech Group), anti‐ANP (Santa Cruz), anti–cMyBP‐C (Santa Cruz), polyclonal anti–cMyBP‐C motif (supplied by C. Witt University of Heidelberg, Heidelberg, Germany), and anti–nuclear factor of activated T cells (NFAT)c4 (Santa Cruz) were used. The isotype‐specific secondary antibodies, Alexa Fluor 488 chicken anti‐rabbit IgG, Alexa Fluor 594 goat anti‐mouse IgG1, Alexa Fluor 488 goat anti‐rat IgM, Alexa Fluor 594 goat anti‐mouse IgM, Alexa Fluor 488 goat anti‐mouse IgG, Alexa Fluor 594 goat anti‐mouse IgG2b, Alexa Fluor 594 chicken anti‐goat IgG, and Alexa Fluor 555 goat anti‐rabbit IgG, were all obtained from Invitrogen. The tested drugs included endothelin‐1 (1.0, 10, 100, or 1000 nmol/L), angiotensin II (100 nmol/L), insulin‐like growth factor 1 (100 nmol/L), phenylephrine (0.05 mmol/L), BQ‐123 (250 nmol/L), and BQ‐788 (100 nmol/L) (all from Sigma).
Tanaka A., Yuasa S., Mearini G., Egashira T., Seki T., Kodaira M., Kusumoto D., Kuroda Y., Okata S., Suzuki T., Inohara T., Arimura T., Makino S., Kimura K., Kimura A., Furukawa T., Carrier L., Node K, & Fukuda K. (2014). Endothelin‐1 Induces Myofibrillar Disarray and Contractile Vector Variability in Hypertrophic Cardiomyopathy–Induced Pluripotent Stem Cell–Derived Cardiomyocytes. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 3(6), e001263.
+ Open protocol
+ Expand
4

Immunohistochemical Analysis of Chronic Pancreatitis

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Human chronic pancreatitis samples were obtained from 9 different patients and were derived from the Institute of Pathology of the Philipps University Marburg in accordance with the ethical regulations of the institute. For immunohistochemistry, formalin-fixed, paraffin-embedded tissue was sectioned (4 μm). H&E staining and immunohistochemistry were performed as described previously [22 (link)]. The following antibodies were used: NFATc4 (Santa Cruz, sc13036), EGFR (Abcam, ab52894), pEGFR (Santa Cruz, sc101668), and Sox9 (Abcam, ab26414).
Hessmann E., Zhang J.S., Chen N.M., Hasselluhn M., Liou G.Y., Storz P., Ellenrieder V., Billadeau D.D, & Koenig A. (2015). NFATc4 Regulates Sox9 Gene Expression in Acinar Cell Plasticity and Pancreatic Cancer Initiation. Stem Cells International, 2016, 5272498.
+ Open protocol
+ Expand
5

ChIP-qPCR Profiling of Transcription Factors

Cited 2 times
Check if the same lab product or an alternative is used in the 5 most similar protocols
ChIP was performed as previously described (Ding et al., 2013 (link)). For tissues, nuclei were first purified by Percoll gradient centrifugation (Wang et al., 2011 (link); Ding and Kilpatrick, 2013a (link)) or differential centrifugation (1400 × g for 3 min, 4°C) in 0.25 M sucrose. Samples were assayed by real-time PCR, and data from triplicate biological replicates were expressed as the fold enrichment relative to antibody controls. ChIP antibodies were to ETV1 (sc-28681; Santa Cruz Biotechnology, Dallas, TX), Xenopus NFIB1, which recognizes mammalian NFIA and NFIB (Ding et al., 2013 (link)), NFIA (39036; Active Motif, Carlsbad, CA), and NFATc4 (sc-1153; Santa Cruz Biotechnology). The term NFI is used to distinguish ChIP assays employing the Xenopus NFIB1 antibody from those specifically assaying NFIA occupancy. Note that the NFI and NFIA antibodies both detect binding to the same genomic regions in ChIP assays (Ding et al., 2013 (link)). Preimmune serum or normal rabbit immunoglobulin G (IgG; PP64; Millipore, Billerica, MA) was used as negative control. PCR primer sequences for ChIP assays are available upon request.
Ding B., Cave J.W., Dobner P.R., Mullikin-Kilpatrick D., Bartzokis M., Zhu H., Chow C.W., Gronostajski R.M, & Kilpatrick D.L. (2016). Reciprocal autoregulation by NFI occupancy and ETV1 promotes the developmental expression of dendrite-synapse genes in cerebellar granule neurons. Molecular Biology of the Cell, 27(9), 1488-1499.
+ Open protocol
+ Expand
6

Chromatin Immunoprecipitation Analysis

Cited 3 times
Check if the same lab product or an alternative is used in the 5 most similar protocols
Nuclei were isolated from fixed tissues and cell cultures using either Percoll gradient centrifugation (Wang et al., 2011 (link); Ding and Kilpatrick, 2013 ) or differential centrifugation (1400 × g for 3 min, 4°C) in 0.25 M sucrose (Ding et al., 2013 (link), 2016 (link)). Chromatin was prepared as described previously (Ding et al., 2013 (link), 2016 (link)), and samples were assayed by real-time PCR as for RT-qPCR analyses. Results from triplicate biological replicates were expressed as the fold enrichment relative to antibody controls. ChIP antibodies were used for ETV1 (sc-28681; Santa Cruz Biotechnology), Xenopus NFIB1, which recognizes mammalian NFIA and NFIB (Ding et al., 2013 (link), 2016 (link)), NFIA (39036; Active Motif), and NFATc4 (sc-1153; Santa Cruz Biotechnology). The term NFI is used to distinguish ChIP assays employing the Xenopus NFIB1 antibody from those specifically assaying NFIA occupancy. Note that the NFI and NFIA antibodies both detect binding to the same genomic regions in ChIP assays (Ding et al., 2013 (link), 2016 (link)). Preimmune serum or normal rabbit immunoglobulin G (IgG [PP64]; EMD Millipore) was used as a negative control antibody. PCR primer sequences for ChIP assays are available upon request.
Ding B., Dobner P.R., Mullikin-Kilpatrick D., Wang W., Zhu H., Chow C.W., Cave J.W., Gronostajski R.M, & Kilpatrick D.L. (2018). BDNF activates an NFI-dependent neurodevelopmental timing program by sequestering NFATc4. Molecular Biology of the Cell, 29(8), 975-987.
+ Open protocol
+ Expand
7

Recombinant M-CSF Protocol for Immunoblotting

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Recombinant mouse M-CSF was purchased from PeproTech (Catalog# 315-02, Rocky Hill, NJ). Antibodies for immunoblotting of proteins were purchased from different companies as indicated: NFATc3, NFATc2, NFATc1-Cell Signaling Technology (Catalog# 4998, 4389, 8032, Danvers, MA), NFATc4–Santa Cruz (sc-13036, Dallas, TX), and α-Tubulin–Santa Cruz (sc-8035). All the antibodies were validated in our earlier publications [6 (link)]. All primers and oligonucleotides were synthesized by IDT (Coralville, IA) and listed in supplement I. ChIP One Day Chromatin Immunoprecipitation kit was purchased from QIAGEN (Catalog# 334471, Germantown, MD).
Karpurapu M., Lee Y.G., Qian Z., Wen J., Ballinger M.N., Rusu L., Chung S., Deng J., Qian F., Reader B.F., Nirujogi T.S., Park G.Y., Pei D, & Christman J.W. (2018). Inhibition of nuclear factor of activated T cells (NFAT) c3 activation attenuates acute lung injury and pulmonary edema in murine models of sepsis. Oncotarget, 9(12), 10606-10620.
+ Open protocol
+ Expand
8

NFAT Translocation Kinetics in LPS-Stimulated BMDM

Check if the same lab product or an alternative is used in the 5 most similar protocols
The BMDM from WT mice were treated with LPS (100 ng/ml) for 0, 0.5, and 4 h and washed in cold 1xPBS. The cellular and nuclear protein fractions were separated by Pierce NE-PER Nuclear and Cytoplasmic Extraction Reagents kit (cat 78833), resolved on 10% SDS-Poly acrylamide gels and immunoblotted with different NFAT antibodies. The antibodies used for immunoblotting were NFATc1 (8032S, Cell Signaling), NFATc2 (4389S, Cell Signaling), NFATc3 (4998S, Cell Signaling) and NFATc4 (sc-13036, SantaCruz Biotechnology, Inc). Purity of cytoplasmic and nuclear preparations was assessed by b-actin and p53 (sc-6243, SantaCruz Biotechnology, Inc).
Ranjan R., Deng J., Chung S., Lee Y.G., Park G.Y., Xiao L., Joo M., Christman J.W, & Karpurapu M. (2014). The Transcription Factor Nuclear Factor of Activated T Cells c3 Modulates the Function of Macrophages in Sepsis. Journal of Innate Immunity, 6(6), 754-764.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!