Anti stat6

Manufactured by Santa Cruz Biotechnology

Sourced in Germany

Anti-STAT6 is a laboratory reagent used in biochemical and molecular biology applications. It is an antibody that specifically binds to the STAT6 protein, which is a transcription factor involved in cellular signaling pathways. The primary function of Anti-STAT6 is to detect and quantify the STAT6 protein in biological samples.

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STAT6 (28 citations) Anti-P-STAT6 (5 citations) Anti-STAT6 (SC-981X (4 citations) Anti-phospho-STAT6 (SC-11762X (2 citations) Anti-STAT6 (M-20, (2 citations) Anti-Stat6 (sc-621; (2 citations)

6 protocols using anti stat6

Immunoblot Analysis of Cellular Proteins

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The primary antibodies purchased from Santa Cruz Biotechnology are specified as follows: anti-STAT6: sc-374021, anti-p-STAT6: sc-136019, anti-Nrf2: sc-13032, anti-NQO1: sc-32793, anti-VDR: sc-13133, anti-CYP24A: sc-365700, anti-Histone H3: sc-517576, anti-Tublin: sc-8035, anti-ATG7: sc-376212, anti-Beclin1: sc-48381, and anti-GAPDH: sc-32233. Antibody against LC3B was purchased from novusbio (NB100-2220). Secondary antibodies conjugated with horseradish peroxidase (HRP) were from Immunoway (Plano, TX: RS0001 (Mouse) and RS0002 (Rabbit)). Human bronchial epithelial cell BEAS-2B and human acute monocytic leukemia cell line (THP-1) were purchased from ATCC (Manassas, VA). BEAS-2B cells were cultured in BEBM with additives from Lonza/Clonetics Corporation (CC-3170), and THP-1 cells were cultured in RPMI 1640 medium supplemented with 10% FBS (Hyclone, Logan, UT) and 0.1% gentamycin (Invitrogen, Carlsbad, CA). For the differentiation of THP-1 cells, 5 ng/ml phorbol-12-myristate-13-acetate (PMA) obtained from Sigma was used. The cells were maintained at 37°C in a humidified incubator containing 5% CO₂.

Yang Y., Li Q., Wei S., Chu K., Xue L., Liu J., Ma Y, & Tao S. (2022). STAT6/VDR Axis Mitigates Lung Inflammatory Injury by Promoting Nrf2 Signaling Pathway. Oxidative Medicine and Cellular Longevity, 2022, 2485250.

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IL4/IL13 Induced STAT6 Activation

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Cells were stimulated with human recombinant IL4 (BD Pharmingen), IL13 (Peprotech) or medium for 4, 8, and 24 hours. STAT6 expression and phosphorylation were determined by Western blot (anti-pSTAT6, BD Biosciences; anti-STAT6 Santa Cruz Biotechnology).

Derks S., Nason K.S., Liao X., Stachler M.D., Liu K.X., Liu J.B., Sicinska E., Goldberg M.S., Freeman G.J., Rodig S.J., Davison J.M, & Bass A.J. (2015). Epithelial PD-L2 expression marks Barrett's Esophagus and Esophageal Adenocarcinoma. Cancer immunology research, 3(10), 1123-1129.

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Chromatin Immunoprecipitation of Transcription Factors

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All antibodies used in these experiments were either conjugated in-house or purchased as indicated. Anti-IL-4 (11B11), anti-IL-12 (C17.18), anti-IFNγ (AN17.18.24), and anti-CD4 (GK1.5) antibodies were purified from hybridoma supernatants at the German Rheumatism Research Center and used at 10 μg/ml final concentration. FITC-conjugated anti-IFNγ (AN18.17.24; BD Pharmingen, Heidelberg, Germany), phycoerythrin-conjugated anti-IL-4 (11B11; BD Pharmingen, and BVD4–1D11; Miltenyi Biotec, Bergisch Gladbach, Germany) were used for intracellular cytokine staining. For the chromatin immunoprecipitation, the following antibodies were used: anti-c-MAF, anti-RNA polymerase II, anti-p300 and anti-STAT6 (polyclonal rabbit IgG; Santa Cruz Biotechnology, Heidelberg, Germany), anti-NFATc2 and -NFATc1 (polyclonal rabbit IgG; ImmunoGlobe Antikörpertechnik GmbH, Himmelstadt, Germany), anti-GATA-3 (mouse monoclonal; Santa Cruz Biotechnology), anti-NF-kB (polyclonal goat IgG; Santa Cruz Biotechnology), anti-Brg1 (rabbit antiserum; Merck-Millipore, Darmstadt, Germany). For the image cytometry, anti-NFATc2 (rabbit monoclonal, clone D4B1, Cell Signaling Technology, Leiden, The Netherlands) and donkey anti-rabbit IgG (coupled to Alexa Fluor 647, Molecular probes A31573, Darmstadt, Germany) were used.

Köck J., Kreher S., Lehmann K., Riedel R., Bardua M., Lischke T., Jargosch M., Haftmann C., Bendfeldt H., Hatam F., Mashreghi M.F., Baumgrass R., Radbruch A, & Chang H.D. (2014). Nuclear Factor of Activated T Cells Regulates the Expression of Interleukin-4 in Th2 Cells in an All-or-none Fashion. The Journal of Biological Chemistry, 289(39), 26752-26761.

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Western Blot Analysis of Lung Tissue

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Lung tissue was lysed by using RIPA buffer (Thermo Fisher Scientific). Equal amounts of proteins were separated by 10% sodium dodecyl sulphate‐polyacrylamide gel electrophoresis and transferred to polyvinylidene fluoride membranes (Bio‐Rad Laboratories). The membrane was blocked with 5% bovine serum albumin (Sigma‐Aldrich) for 90 min at room temperature and incubated with a primary antibody for 14 h at 4℃. The primary antibodies included anti–PPAR‐γ (Abcam), anti‐TGF‐β1 (ProteinTech), anti‐phosphor‐Smad3 (Cell Signaling Technology), anti‐Smad3 (Cell Signaling Technology), anti‐phosphor‐Stat6 (Cell Signaling Technology), anti‐Stat6 (Santa Cruz Biotechnology), anti‐phosphor‐Stat3 (Cell Signaling Technology), anti‐Stat3 (Santa Cruz Biotechnology), anti‐β‐tubulin (ProteinTech), anti‐NFκB P65 (Abcam), anti‐IκBα (Abcam), anti‐IL‐1β (Abcam), anti‐phosphor‐NFκB P65 (Cell Signaling Technology), anti‐phosphor‐IκBα (Cell Signaling Technology) and anti‐GAPDH (Abcam). And then, membranes were incubated with anti‐rabbit/mouse IgG (H+L) (Abcam) for 1 h at room temperature. Chemiluminescence measurement was performed by using Amersham Imager 680 (GE Healthcare Life Science).

Wang S., Chen Y., Hong W., Li B., Zhou Y, & Ran P. (2022). Chronic exposure to biomass ambient particulate matter triggers alveolar macrophage polarization and activation in the rat lung. Journal of Cellular and Molecular Medicine, 26(4), 1156-1168.

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STAT1-Dependent Cytokine Signaling in Parasitic Infection

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Peritoneal exudate cells from STAT1^+/+ and STAT1^−/− mice infected with Taenia crassiceps and controls were incubated for 24 h; subsequently, the adherent cells were washed with culture medium and stimulated with or without IL-4 recombinant (20 ng/mL; Peprotech Mexico) for 20 min. Cells were lysed and then 30 µg of protein was boiled for 5 min and separated by SDS-PAGE using 10% polyacrylamide gel, and transferred to polyvinylidene difluoride (PVDF) membranes (Immobilon-P, Millipore). Membranes were blocked with 5% fat-free milk in PBS for 1 h at room temperature and incubated overnight at 4 °C with the primary antibody. The primary antibodies were recognized (Anti-β-actin Biolegend,1:2000; Anti-AKT Cell Signalling,1:1000; Anti-AKT-P Cell Signalling,1:1000; Anti-STAT6 Santa Cruz,1:200, Anti-STAT6-P Santa Cruz,1:150). Then, the detection step was performed with HRP-coupled anti-rabbit IgG (BioLegend 1:2000). Finally, the detection was done with the Chemiluminescent Substrate Kit (VisiGlo, VWR, AMRESCO) and the bands were visualized with the C-DiGit (LI-COR) equipment.

Becerra-Díaz M., Ledesma-Soto Y., Olguín J.E., Sánchez-Barrera A., Mendoza-Rodríguez M.G., Reyes S., Satoskar A.R, & Terrazas L.I. (2021). STAT1-Dependent Recruitment of Ly6ChiCCR2+ Inflammatory Monocytes and M2 Macrophages in a Helminth Infection. Pathogens, 10(10), 1287.

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ChIP-qPCR Analysis of Histone Modifications and Transcription Factors

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Upon protein-DNA cross-linking, cells were lysed and sonicated to achieve chromatin fragments ranging between 200 and 1000 bp in size. The lysates were incubated with anti-H3Ac (Millipore), anti-STAT5 (Cell Signaling Technologies), anti-STAT6 (Santa Cruz Biotechnology) or isotype controls, and then complexes were isolated using Dynabeads Protein A (Invitrogen). Immunoprecipitates were extensively washed and then eluted with freshly prepared 1% SDS, 0.1 M NaHCO3 buffer. After reversion of cross-linking and deproteinization, DNA was purified with QIAquick PCR purification kit (Qiagen) followed by real time PCR analysis. The following primer pairs were used: R1 forward 5’-cagctgtgcgtggaggcttttca-3’; R1 reverse 5’-tccttccctagaccagtgaagttcgaaga-3’; R2 forward 5’-tgcaccagccttgaactgaaccag-3’; R2 reverse 5’-gctacacaactgcaagggacagctgatta-3’; R3 forward 5’-ccaccccagcctatagtgag-3’; R3 reverse 5’-tagtctccagctggctcagg-3’; R4 forward 5’-cgttctgaacacgggagat-3’; R4 reverse 5’-tgggtgagctcatgtctctg-3’; R5 forward 5’-gcctaccatgtgccgagata-3’; R5 reverse 5’-agattacatctttcagtgaagtcaa-3’; Nuc1 forward 5’-gcctaccatgtgccgagata-3’; Nuc1 reverse 5’-tgtttactatctgtctcccacctg-3’.

Borriello F., Longo M., Spinelli R., Pecoraro A., Granata F., Staiano R.I., Loffredo S., Spadaro G., Beguinot F., Schroeder J, & Marone G. (2015). IL-3 synergizes with basophil-derived IL-4 and IL-13 to promote the alternative activation of human monocytes. European journal of immunology, 45(7), 2042-2051.

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