Anti stat4

Manufactured by Santa Cruz Biotechnology

Anti-STAT4 is a primary antibody that targets the STAT4 protein. STAT4 is a transcription factor that plays a key role in the signaling pathways of certain cytokines, such as interleukin-12 (IL-12) and type I interferons. This antibody can be used for various applications, including Western blotting, immunoprecipitation, and immunohistochemistry, to detect and study the STAT4 protein in biological samples.

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Lab products found in correlation

Anti stat3, by Santa Cruz Biotechnology (3 mentions) Anti erk, by Cell Signaling Technology (1 mentions) Anti p akt, by Cell Signaling Technology (1 mentions) Anti p erk, by Cell Signaling Technology (1 mentions) Anti akt, by Santa Cruz Biotechnology (1 mentions) Anti pparα, by Abcam (1 mentions) Anti acly, by Santa Cruz Biotechnology (1 mentions) Anti fasn, by Santa Cruz Biotechnology (1 mentions) Anti pstat4, by Santa Cruz Biotechnology (1 mentions) Anti pmapk, by Cell Signaling Technology (1 mentions) Anti β actin, by Merck Group (1 mentions) Anti h3k27ac, by Abcam (1 mentions) Protein g dynabead, by Thermo Fisher Scientific (1 mentions) Sybr select master mix, by Thermo Fisher Scientific (1 mentions) Chemiluminescence reagent, by Thermo Fisher Scientific (1 mentions) Hrp conjugated goat anti rabbit igg secondary antibody, by Merck Group (1 mentions) Laminin b, by Abcam (1 mentions) Anti gr, by Abcam (1 mentions) Anti c jun, by Abcam (1 mentions) Anti nf κb p65, by Abcam (1 mentions)

Close spelling variants of this product

Anti-STAT4 (sc486; STAT4

5 protocols using anti stat4

Western Blot Antibody Characterization

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Standard Western-blot assays were used as described previously^{59 (link)}. The following primary antibodies were used: anti-PPARα (Abcam, Cata# ab24509), anti-ACLY (Santa Cruz, Cata# sc-517267), anti-FASN (Santa Cruz, Cata# sc-48357), anti-ACSL1 (cell signaling, Cata# 4047 S), anti-ACSL5 (Santa Cruz, Cata# sc-365478), anti-STAT3 (Santa Cruz, Cata# sc-8019), anti-pSTAT3 (cell signaling, Cata# 9145 S), anti-STAT1 (cell signaling, Cata# 14994 S), anti-pSTAT1 (cell signaling, Cata# 9177 S), anti-STAT4 (cell signaling, Cata# 2653 S), anti-pSTAT4 (cell signaling, Cata# 4134 S), anti-AKT (Santa Cruz, Cata# sc-5298, 1:2000 dilution), anti-pAKT (cell signaling, Cata# 9018 S), anti-pERK (cell signaling, Cata# 4376), anti-ERK (cell signaling, Cata# 9102), anti-pMAPK (cell signaling, Cata# 4511), anti-MAPK (cell signaling, Cata# 9212) and anti-β-actin (Sigma, Cata# A5441, 1:100,000 dilution) antibodies. Other than anti-AKT and anti-β-actin antibodies, the dilution for all other antibodies was 1:1000.

Yang X., Wang J., Chang C.Y., Zhou F., Liu J., Xu H., Ibrahim M., Gomez M., Guo G.L., Liu H., Zong W.X., Wondisford F.E., Su X., White E., Feng Z, & Hu W. (2024). Leukemia inhibitory factor suppresses hepatic de novo lipogenesis and induces cachexia in mice. Nature Communications, 15, 627.

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ChIP-qPCR Analysis of STAT4 and H3K27Ac

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Approximately 5 × 10⁶ enriched splenic NK cells were purified by sorting on TCRβ⁻NK1.1⁺ cells. Proteins were cross-linked to chromatinized DNA for 8 min at 25 °C by addition of 1% formaldehyde to the medium. ChIP was carried out as previously described^{63 (link)} using 10 μg of rabbit polyclonal anti-STAT4 (Santa Cruz, sc-468, clone C-20) or 10 μg of rabbit serum IgG antibody as a control (data not shown; Sigma, I5006), or 0.5 μg of anti-H3K27Ac (Abcam, 4729). Relative abundance of regulatory sequences in the Zbtb32 promoter or, as controls, in the Actb and Gapdh promoters or a gene desert ∼50 kb upstream of the Foxp3 gene was measured by qPCR in the antibody-precipitated DNA. Percent input was calculated by 100 × 2̂(CT^{adjusted input} – CT^target), where the CT^input was adjusted from 5% to 100% by subtracting Log₂ 20 CTs. Primer sequences are listed in Supplementary Table 2.

Beaulieu A.M., Zawislak C.L., Nakayama T, & Sun J.C. (2014). The transcription factor Zbtb32 controls the proliferative burst of virus-specific natural killer cells responding to infection. Nature immunology, 15(6), 546-553.

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ChIP Assay for STAT3 and RORγt Transcription Factors

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The ChIP procedure was performed using an assay kit following the manufacturer's instruction (EMD Millipore). Briefly, T_H17 and T_H1 cells were cross-linked by 1% formaldehyde for 10 min at 37^°C. Nuclei were prepared and subjected to sonication to obtain DNA fragments. Chromatin fractions were precleared with protein A-agarose beads followed by immunoprecipitation overnight at 4^°C with 3 μg of anti-STAT3 (Santa Cruz), anti-ROR gamma(t) (eBioscience), anti-STAT4 (Santa Cruz), or control antibody. Cross-linking was reversed at 65^°C for 4 hrs, followed by proteinase K digestion. DNA was purified and subjected to qPCR. The input DNA was diluted 200 times prior to PCR amplification. The input and immunoprecipitated DNA were amplified by qPCR using primers encompassing the STAT3 or STAT4 binding sites of the mouse RORγt or T-bet promoter regions.

Wang J., Peng L., Zhang R., Zheng Z., Chen C., Cheung K.L., Cui M., Bian G., Xu F., Chiang D., Hu Y., Chen Y., Lu G., Yang J., Zhang H., Yang J., Zhu H., Chen S.H., Liu K., Zhou M.M., Sikora A.G., Li L., Jiang B, & Xiong H. (2016). 5-Fluorouracil targets thymidylate synthase in the selective suppression of TH17 cell differentiation. Oncotarget, 7(15), 19312-19326.

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Chromatin Immunoprecipitation for Transcription Factors

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Chromatin was prepared from the indicated T helper cell population as previously described^{17 (link)}. The resulting chromatin was incubated with either anti-STAT4 (Santa Cruz), anti-STAT3 (Santa Cruz), or control antibody (Abcam) and immunoprecipitated using Protein G Dynabeads (Life Technologies). Precipitated DNA was analyzed via quantitative PCR using SYBR Select Mastermix (Life Technologies) and gene-specific primers: (Bcl6 promoter forward: 5′-GCGGAGCAATGGTAA AGCCC-3′, and reverse: 5′-CTGGTGTCCGGCCTTTCCTAG-3′; Bcl6 control forward: 5′-GTACTCCAACAACAGCACAGC-3′, and reverse: 5′-GTGGCTCGTTAAATCACAGAGG-3′; Il21 promoter forward: 5′-CAC ACACCTTGGTGAATGCTG-3′, and reverse: 5′-CCATTGGCTAGGTGTACGTGTG-3′). Samples were normalized to total input followed by the subtraction of the isotype control to account for unspecific binding.

Powell M.D., Read K.A., Sreekumar B.K., Jones D.M, & Oestreich K.J. (2019). IL-12 signaling drives the differentiation and function of a TH1-derived TFH1-like cell population. Scientific Reports, 9, 13991.

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Western Blot Analysis of NK92 Cells

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For Western blot analysis, nuclear and cytoplasmic compartments of NK92 cells were extracted from 1–3 × 10⁶ cells via the Fermentas ProteoJET Cytoplasmic and Nuclear Protein Extraction protocol (Fermentas, Burlington, ON). Nuclei were lysed using Nuclear Lysis Buffer (Fermentas, Burlington, ON) and both lysed nuclei and cytoplasm were resuspended in Laemmlis SDS-Sample Buffer (4×) (Boston Bioproducts, Boston, MA). Samples were boiled for 10 minutes and proteins separated by electrophoresis with a 12% agarose gel and transferred to a nitrocellulose membrane for blotting. Proteins were visualized with anti-NF-κB p65 (Abcam, Cambridge, MA), anti-cJun (Abcam, Cambridge, MA), anti-STAT4 (Santa Cruz, Santa Cruz, CA), anti-GR (Abcam, Cambridge, MA) and HRP conjugated goat anti-rabbit IgG secondary antibody (Millipore, Temecula, CA) and chemiluminescence reagent (ThermoScientific, Rockford, IL). Quality of nuclear and cytoplasmic compartment extraction was determined by Laminin B and GAPDH, respectively (Abcam, Cambridge, MA). Blot density was quantified using Image J software.

Eddy J.L., Krukowski K., Janusek L, & Mathews H.L. (2014). Glucocorticoids regulate natural killer cell function epigenetically. Cellular immunology, 290(1), 120-130.

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