Rabbit anti stat6

Manufactured by Cell Signaling Technology

Rabbit anti-STAT6 is a primary antibody that recognizes the STAT6 protein. STAT6 is a transcription factor that plays a key role in the IL-4 and IL-13 signaling pathways. This antibody can be used for applications such as Western blotting and immunoprecipitation to detect and study the STAT6 protein.

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STAT6 (53 citations) Anti-STAT6 (30 citations) Rabbit anti-phospho-STAT6 (5 citations)

4 protocols using rabbit anti stat6

Protein Interaction Analysis by Co-Immunoprecipitation

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Co-immunoprecipitations and immunoblot analyses were performed as previously described (70 (link)). Subcellular fractionation for co-immunoprecipitations on chromatin fractions was performed using the Subcellular Protein Fractionation Kit (Thermo Scientific, #78840) according to the manufacturer’s instructions. The primary antibodies used were as follows: mouse anti-p21 (Santa Cruz, sc-53870; 1:8000 used for both mouse and human samples), rabbit anti-Myc-tag (Cell Signaling, #2272; 1:1000); rabbit anti-Rb (Abcam, ab181616; 1:2000), rabbit anti-STAT1 (Abcam, ab92506; 1:1000), rabbit anti-STAT6 (Cell Signaling, #5397; 1:1000), rabbit anti-SMAD2 (Cell Signaling, #5339, 1:1000), rabbit anti-SMAD3 (Cell Signaling, #9513; 1:1000), and mouse anti-p27 (BD Biosciences, #610242, 1:1000). All antibodies were detected with secondary HRP-conjugated goat anti-mouse or anti-rabbit antibodies (Jackson Immunoresearch; 1:10,000). PonS staining (0.2% w/v in 5% glacial acetic acid, Sigma-Aldrich, #P3504) served as a loading control. Immunoblots are representative of at least two independent experiments.

Sturmlechner I., Zhang C., Sine C.C., van Deursen E.J., Jeganathan K.B., Hamada N., Grasic J., Friedman D., Stutchman J.T., Can I., Hamada M., Lim D.Y., Lee J.H., Ordog T., Laberge R.M., Shapiro V., Baker D.J., Li H, & van Deursen J.M. (2021). p21 produces a bioactive secretome that places stressed cells under immunosurveillance. Science (New York, N.Y.), 374(6567), eabb3420.

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Western Blot Analysis of Cellular Signaling Pathways

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For Western blotting, 300,000 MN9D cells were grown overnight in 35 mm
dishes prior to the indicated treatments. Total protein extracts were prepared
and analyzed by SDS-PAGE and Western blotting as described previously
[5 (link)]. The primary
antibodies used for Western blotting were: rabbit anti-phospho-Akt (ser473)
(#9271, 1/1000), rabbit anti-phospho-Stat6 (#9361, 1/1000),
rabbit anti-phospho-4E-BP1 (thr37/46) (#2855, 1/10,000), rabbit anti-Akt
(#9272, 1/10,000), rabbit anti-Stat6 (#9362, 1/1000), rabbit
anti-4E-BP1 (#9644, 1/250,000) and HRP-conjugated rabbit anti-actin
(#5125, 1/20,000) from Cell Signaling. Following washing, the Western
blots were incubated for 1 hr at room temperature in horseradish peroxidase-goat
anti-rabbit or goat anti-mouse (Biorad, Hercules, CA) diluted 1/2500 and
developed with the Super Signal reagent (Pierce, Rockford, IL). In all cases,
the same membrane was re-probed for actin and/or a parallel membrane was probed
with an antibody reacting with the total protein in order to provide a
normalization standard. Autoradiographs were scanned and analyzed using a Biorad
GS800 scanner. The experiments were repeated a minimum of three times with
independent protein samples.

Maher P, & Conti B. (2017). Deciphering the Pathways that Protect from IL-13-Mediated Potentiation of Oxidative Stress-Induced Dopaminergic Nerve Cell Death. Cytokine, 103, 114-120.

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Western Blot Analysis of Inflammatory Markers

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Proteins from BMDMs and ipsilateral cortical tissue were extracted using RIPA buffer, equalized, and loaded onto 5–20% gradient gels for SDS PAGE (Bio-Rad; Hercules, CA). Proteins were transferred onto nitrocellulose membranes and then blocked for 1 h in 5% milk in 1 × TBS containing 0.05% Tween-20 (TBS-T) at room temperature. The membrane was incubated in mouse anti-arginase 1 (N-20) (1:1000; BD Transduction Laboratories, San Jose, CA), rabbit anti-STAT6 (1:1000; Cell signaling, Danvers, MA), rabbit anti-pSTAT6 (1:1000; Cell signaling), mouse anti-STAT3 (1:1000; Cell signaling), rabbit anti-pSTAT3 (1:1000; Cell signaling), or mouse anti-β-Actin (1:5000; Sigma-Aldrich) overnight at 4 °C, then washed three times in TBS-T, and incubated in appropriate HRP-conjugated secondary antibodies (Jackson ImmunoResearch Laboratories, West Grove, PA) for 2 h at room temperature. Membranes were washed three times in TBS-T, and proteins were visualized using SuperSignal West Dura Extended Duration Substrate (Thermo Scientific, Rockford, IL). Chemiluminescence was captured ChemiDoc™ XRS+ System (Bio-Rad), and protein bands were quantified by densitometric analysis using BioRad Molecular Imaging Software. The data presented reflects the intensity of target protein band normalized based on the intensity of the endogenous control for each sample (expressed in arbitrary units).

Barrett J.P., Henry R.J., Villapol S., Stoica B.A., Kumar A., Burns M.P., Faden A.I, & Loane D.J. (2017). NOX2 deficiency alters macrophage phenotype through an IL-10/STAT3 dependent mechanism: implications for traumatic brain injury. Journal of Neuroinflammation, 14, 65.

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Western Blot Analysis of Post-SAH Brain

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At each post-SAH time point, the brain samples were collected and prepared for western blot analysis. The left hemisphere sample was prepared using RIPA lysis buffer. 40 ng of protein sample was loaded onto an SDS-PAGE gel and transferred to a nitrocellulose membrane. The membrane was blocked with 5% non-fat milk and incubated with the following primary antibodies: rabbit anti-CCR4 (1:1000, GTX53474, Gene Tex); rabbit anti-CCL17 (1:1000, ab182793, Abcam); rabbit anti-Akt (1:1000, 9272S, Cell Signaling Technology); rabbit anti-phosphorylated AKT ser473 (1:1000, 4060S, Cell Signaling Technology); rabbit anti-PKCα (1:000, 2056S, Cell Signaling Technology); rabbit antiphosphorylated PKCα (1:1000, ab180848, Abcam); rabbit anti-PKC (1:1000, (ab179522, Abcam); rabbit anti-phosphorylated 4EBP1 (1:1000, 2855S, Cell Signaling Technology), rabbit anti-4EBP1 (1:1000, 9644S, Cell Signaling Technology); rabbit anti-phosphorylated STAT6 (1:1000, 5397S, Cell Signaling Technology); rabbit anti-STAT6 (1:1000, 56554S, Cell Signaling Technology); and rabbit anti-β-actin (1:5000, 3700S, Cell Signaling Technology) overnight at 4 °C. Membranes were incubated with corresponding secondary antibody (1: 10000, Santa Cruz; 1:5000, Abcam) for 2 h at room temperature.

Zhang A., Liu Y., Xu H., Zhang Z., Wang X., Wang L., Lenahan C., Zhang C., Jiang J., Fang C., Fang Y., Zhang J., & Chen S. (2022). Neuron-derived CCL17 Exerts Neuroprotection Through Activation of CCR4/mTORC2 Axis in Microglia After Subarachnoid Hemorrhage in Rats.

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