Log In Sign up
The largest database of trusted experimental protocols

Phosphorylated stat3

Manufactured by Cell Signaling Technology
Visit Supplier
Sourced in United States, Germany

Phosphorylated STAT3 is a laboratory product that detects the phosphorylated form of the STAT3 protein. STAT3 (Signal Transducer and Activator of Transcription 3) is a transcription factor that plays a key role in cellular signaling pathways.

Automatically generated - may contain errors

Lab products found in correlation

Stat3, by Cell Signaling Technology (16 mentions) Phosphorylated akt, by Cell Signaling Technology (10 mentions) Total stat3, by Cell Signaling Technology (8 mentions) Phosphorylated erk, by Cell Signaling Technology (7 mentions) β actin, by Santa Cruz Biotechnology (5 mentions) Cleaved caspase 3, by Cell Signaling Technology (5 mentions) β actin, by Merck Group (4 mentions) β actin, by Cell Signaling Technology (4 mentions) Erk1 2, by Cell Signaling Technology (3 mentions) Phosphorylated stat1, by Cell Signaling Technology (3 mentions) Cleaved parp, by Cell Signaling Technology (3 mentions) E cadherin, by Cell Signaling Technology (3 mentions) Vimentin, by Cell Signaling Technology (3 mentions) Proliferating cell nuclear antigen (pcna), by Cell Signaling Technology (2 mentions) Phosphorylated iκbα, by Cell Signaling Technology (2 mentions) α sma, by Abcam (2 mentions) Cox 2, by Santa Cruz Biotechnology (2 mentions) Igf 1r, by Cell Signaling Technology (2 mentions) Proliferating cell nuclear antigen (pcna), by Santa Cruz Biotechnology (2 mentions) Cd11b, by Thermo Fisher Scientific (2 mentions)

Close spelling variants of this product

STAT3 (1075 citations) Anti-phosphorylated STAT3 (20 citations) Phosphorylated STAT3 (Tyr705), (9 citations) Phosphorylated STAT3 (p-STAT3) (6 citations) Rabbit anti-phosphorylated STAT3 (5 citations) Anti-phosphorylated STAT3 (Tyr705 (5 citations) Anti-phosphorylated STAT3 antibody (4 citations) Anti-phosphorylated STAT3 (p-STAT3) (4 citations) Anti-phosphorylated (p-)STAT3 ( (3 citations) Rabbit monoclonal anti-phosphorylated STAT3 (3 citations)

50 protocols using phosphorylated stat3

1

Western Blot Protein Expression Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
A standard western blot assay was used to analyse protein expression, using primary antibodies against signal transducer and activator of transcription 3 (STAT3) (1 : 1000; Cell Signaling Technology, Beverly, Massachusetts, USA), phosphorylated STAT3 (1 : 1000; Cell Signaling Technology), extracellular signal‐regulated kinase (ERK) 1 (1 : 1000; Santa Cruz Biotechnology, Santa Cruz, USA), phosphorylated ERK (1 : 2000; Cell Signaling Technology), PCNA (1 : 1000; Cell Signaling Technology) and β‐tubulin (1 : 5000; Sigma Aldrich), as described previously20. The immunoreactive signals were visualized by scanning densitometry with the ChemiDoc™ Touch Imaging System (Bio‐Rad Laboratories, Hercules, California, USA).
Shi J.H., Yan X., Zhang S.J, & Line P.D. (2020). Simulated model of RAPID concept: highlighting innate inflammation and liver regeneration. BJS Open, 4(5), 893-903.
+ Open protocol
+ Expand
2

Histological Analysis of Knee Joints

Check if the same lab product or an alternative is used in the 5 most similar protocols
Knee joints were harvested and fixed in 10% neutral buffered formalin for 3 days, decalcified in Formic Acid Bone Decalcifier (Immunocal, Decal Chemical Corp.) for 7–10 days, paraffin processed, and embedded for sectioning. Tissues were sectioned at 5 µm and stained with ABH/OG. IHC analyses were performed on sections using traditional antigen retrieval and colorimetric development methodologies with the following primary antibodies: SOX9 (Santa Cruz), COL2A1 (Thermo Scientific), COL10A1 (Quartett), COL1A1 (Abcam), COL3A1 (Abcam), MMP-13 (Thermo Scientific), IL6 (Abcam), and phosphorylated STAT3 (Cell Signaling). TUNEL cell death assay was performed on sections using the in situ Cell Death Detection Kit, Fluorescein (Roche) according to the manufacturer’s instructions. IF analysis and β-galactosidase staining was performed on frozen sections. Knee joints were harvested and fixed in 4% paraformaldehyde (PFA) for 2 hours at 4 °C and decalcified with 14% EDTA at 4 °C for 10 days. Tissues were washed in sucrose gradient, embedded with Tissue-TEK OCT medium, snap frozen in liquid nitrogen and sectioned at 10 µm using a Lecia CM 1850 cryotome. The NOTCH1 primary antibody (Santa Cruz) was used for IF analysis. Beta-galactosidase staining was performed as previously described (63 (link)).
Liu Z., Chen J., Mirando A., Wang C., Zuscik M.J., O’Keefe R.J, & Hilton M.J. (2015). A Dual Role for NOTCH Signaling in Joint Cartilage Maintenance and Osteoarthritis. Science signaling, 8(386), ra71.
+ Open protocol
+ Expand
3

Western Blot Analysis of IL-10R1 and STAT3

Check if the same lab product or an alternative is used in the 5 most similar protocols
NRCM were lyzed in 100 µl RIPA buffer (containing 1% IGEPAL CA-630, 0.5% sodium deoxycholate, 0.1% SDS, 0.5 mM phenylmethylsulphonyl fluoride, 500 ng/ml Leupeptin, 1 mg/ml Aprotinin and 2.5 mg/ml Pepstatin A). Western blot analysis was conducted using standard protocols. Proteins were separated by SDS-PAGE and transferred to PVDF membranes (Millipore). Membranes were blocked in 3% BSA and incubated with primary antibodies against IL-10R1 (Santa Cruz), phosphorylated STAT3 or total STAT3 (Cell Signaling) overnight, followed by HRP-linked anti-rabbit secondary antibody (Cell Signaling) for 2 h. Proteins were visualized using enhanced chemiluminescence (GE Healthcare) on a ChemiDoc XRS Imaging System (Biorad). Membranes were then incubated with β-actin or GAPDH antibody (abcam) as loading control.
Stafford N., Assrafally F., Prehar S., Zi M., De Morais A.M., Maqsood A., Cartwright E.J., Mueller W, & Oceandy D. (2020). Signaling via the Interleukin-10 Receptor Attenuates Cardiac Hypertrophy in Mice During Pressure Overload, but not Isoproterenol Infusion. Frontiers in Pharmacology, 11, 559220.
+ Open protocol
+ Expand
4

Macrophage Inflammatory Signaling Modulation

Check if the same lab product or an alternative is used in the 5 most similar protocols
RAW 264.7 macrophages were plated in 6-well plates (4 × 105/well) and cultured in 2 mL of DMEM for 4 h. The cultures were washed to remove non-adherent cells and then incubated with 2 mL of complete DMEM for 20 h. The culture medium was replaced with DMEM for 30 min to allow the cells to adjust. (i) To induce an inflammation model, 1 μg/mL LPS (Sigma) was added. After 24 h of stimulation with LPS, the cells were treated with the indicated concentrations of L-4F (0, 0.1, or 0.25 μg/mL) for 12 h, and the levels of pp38, pJNK and pERK were analyzed. (ii) Cells were stimulated by LPS and treated with the indicated concentrations of L-4F (0, 0.1, or 0.25 μg/mL) for 1 h, and the levels of pSTAT3 were analyzed.
Western blot procedure: Briefly, cells were lysed in radio-immunoprecipitation assay buffer containing the phosphatase and protease inhibitors phenylmethanesulfonyl fluoride and aprotinin (Sigma), and protein was collected. Then, the protein was separated by SDS-PAGE, transferred to PVDF membranes (Roche) and probed with the indicated primary antibodies (phosphorylated STAT3, phosphorylated p38, phosphorylated JNK and phosphorylated ERK; Cell Signaling Technology). The antibody-antigen complexes were detected using a Chemiluminescent HRP substrate kit (Millipore) according to the manufacturer’s protocols.
Peng M., Zhang Q., Cheng Y., Fu S., Yang H., Guo X., Zhang J., Wang L., Zhang L., Xue Z., Li Y., Da Y., Yao Z., Qiao L, & Zhang R. (2017). Apolipoprotein A-I mimetic peptide 4F suppresses tumor-associated macrophages and pancreatic cancer progression. Oncotarget, 8(59), 99693-99706.
+ Open protocol
+ Expand
5

Keratinocyte and Kidney Cell Culture

Check if the same lab product or an alternative is used in the 5 most similar protocols
Primary keratinocytes were cultured in keratinocyte serum-free media (K-SFM) with supplement purchased from Life Technologies with 100 µg/ml penicillin G and 100 µg/ml streptomycin. Media was changed every 2 d. Kidney epithelial cells were maintained in DMEM supplemented with 10% (volume/volume) FBS (Hyclone), 100 µg/ml penicillin G, and 100 µg/ml streptomycin. Recombinant IL-17A, IL-17F, and TNF were from R&D Systems; anti-Flag (M2) and anti–human STEAP4 antibody was from Sigma-Aldrich; antibody to phosphorylated Jnk, antibody to phosphorylated IκBα, phosphorylated p-38, phosphorylated ERK5, ERK5, phosphorylated STAT3, STAT3, ERK1/2, phosphorylated 4EBP, phosphorylated S6, and anti-p63 for Western blotting were from Cell Signaling Technology; antibody to phosphorylated ERK, actin, anti-BrdU, and anti-p63 for staining were from Santa Cruz Biotechnology, Inc. MEK5 inhibitor (MEK5i, Bix 02189) was ordered from Selleck. IL-6 neutralizing antibody was purchased from BD.
Wu L., Chen X., Zhao J., Martin B., Zepp J.A., Ko J.S., Gu C., Cai G., Ouyang W., Sen G., Stark G.R., Su B., Vines C.M., Tournier C., Hamilton T.A., Vidimos A., Gastman B., Liu C, & Li X. (2015). A novel IL-17 signaling pathway controlling keratinocyte proliferation and tumorigenesis via the TRAF4–ERK5 axis. The Journal of Experimental Medicine, 212(10), 1571-1587.
+ Open protocol
+ Expand
6

Investigating the Anti-Cancer Effects of α-Solanine

Check if the same lab product or an alternative is used in the 5 most similar protocols
α-solanine, dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich(St. Louis, MO, U.S.A.). Protein assay kit was obtained from Bio-Rad Labs (Hercules, CA, U.S.A.). Dulbecco’s modified Eagle’s medium (DMEM) and was purchased from Gibco/BRL (Gaithersburg, MD, U.S.A.). Matrigel was from BD Biosciences (Bedford, MA, U.S.A.). Total RNA extraction kit and polymerase chain reaction (PCR) kit were from Viogene (Sunnyvale, CA, U.S.A.). Antibodies against MMP-2, MMP-9, E-cadherin, TCF-1, STAT3 and phosphorylated STAT3 were purchased from Cell Signaling Technology (Danvers,MA, U.S.A.). Antibodies against β-actin, VEGF, PCNA, β-catenin, Akt, mTOR, phosphorylated proteins were purchased from Abcam(Cambridge, MA, USA). PANC-1, sw1990, MIA PaCa-2 and Human umbilical vein endothelial cell(HUVEC) were obtained from ATCC (Manassas, VA, USA). Pancreatic cancer cell lines are maintained in DMEM with 10% FBS and incubated in a 5% CO2 humidified incubator at 37°C, HUVECs were cultured in M199 medium with 10% FBS. α-Solanine was melted in DMSO and diluted with culture medium (the final concentration of DMSO was less than 0.1%).
Lv C., Kong H., Dong G., Liu L., Tong K., Sun H., Chen B., Zhang C, & Zhou M. (2014). Antitumor Efficacy of α-Solanine against Pancreatic Cancer In Vitro and In Vivo. PLoS ONE, 9(2), e87868.
+ Open protocol
+ Expand
7

Immunoblot and Immunoprecipitation Analysis of Liver Tissues

Check if the same lab product or an alternative is used in the 5 most similar protocols
Liver tissues of mice were lysed in NET-N buffer (20 mM Tris-HCl [pH 8.0], 100 mM NaCl, 1 mM EDTA, 0.5% NP-40) containing protease inhibitor cocktail set I (Merck Millipore, Darmstadt, Germany), and lysates were subjected to immunoblot analysis. For immunoprecipitation, cell lysates were incubated with an anti-TRAF6 antibody (Poteintech, Rosemont, IL) and protein A-Sepharose beads (GE Healthcare, Uppsala, Sweden) overnight and then washed vigorously three times with lysis buffer. Antibodies against STAT3, phosphorylated STAT3, extracellular signal-regulated kinase (ERK), phosphorylated ERK, IκB-α, phosphorylated IκB-α, and phosphorylated IFN regulatory factor (IRF)-7 were purchased from Cell Signaling Technologies (Danvers, MA). Anti-IRAK1 and anti-IRF-7 antibodies were purchase from Santa Cruz Biotechnology (Dallas, TX), and anti-vinculin and anti-glyceraldehyde-3-phosphatate dehydrogenase (GAPDH) antibodies were purchased from Merck Millipore. For immunohistochemistry, liver tissue sections were incubated with an anti-phospho-STAT3 antibody followed by detection using HISTOFINE SimpleStain Mouse MAX-PO(R) (Nichirei Biosciences Inc., Tokyo, Japan).
Tsutsumi T., Okushin K., Enooku K., Fujinaga H., Moriya K., Yotsuyanagi H., Aizaki H., Suzuki T., Matsuura Y, & Koike K. (2017). Nonstructural 5A Protein of Hepatitis C Virus Interferes with Toll-Like Receptor Signaling and Suppresses the Interferon Response in Mouse Liver. PLoS ONE, 12(1), e0170461.
+ Open protocol
+ Expand
8

Analyzing IL22 Signaling in Pancreatitis

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Primary PSCs were treated with murine rIL22 and homogenized in RIPA buffer containing protein inhibitors and analyzed by Western blot, as described previously.16 (link) Antibodies to IL22RA1, α–smooth muscle actin (αSMA; Abcam, Cambridge, MA), signal transducers and activators of transcription (STAT) 3, and phosphorylated STAT3 (Cell Signaling Technologies, Danvers, MA) were used. Antibodies to CYP1a1 (Novus Biologicals, Littleton, CO), β-tubulin (Cell Signaling Technologies), and actin (Sigma) were used in blots of pancreas homogenate from air- or cigarette smoke–exposed mice. Immunofluorescence staining for IL22RA1 and αSMA was performed according to manufacturer’s guideline and analyzed using confocal microscope. Serum samples from healthy subjects and CP patients were analyzed using enzyme-linked immunosorbent assay kit for human IL22 detection (eBioscience).
Xue J., Zhao Q., Sharma V., Nguyen L.P., Lee Y.N., Pham K.L., Edderkaoui M., Pandol S.J., Park W, & Habtezion A. (2016). Aryl Hydrocarbon Receptor Ligands in Cigarette Smoke Induce Production of Interleukin-22 to Promote Pancreatic Fibrosis in Models of Chronic Pancreatitis. Gastroenterology, 151(6), 1206-1217.
+ Open protocol
+ Expand
9

Histopathological Analysis of Tissue Samples

Check if the same lab product or an alternative is used in the 5 most similar protocols
Paraffin-embedded or frozen tissues were stained with H&E, Sirius red, or processed for IHC. Antigens were retrieved with preheated 1xCitrate buffer and were detected by staining with Ki-67 (Genetex, Irvine, CA, USA), cleaved-caspase 3, phosphorylated-STAT3, IGF-1R (Cell Signaling), COX-2, IRS1, PCNA (Santa Cruz), CD11b, GP38 (eBioscience), α-SMA (Abcam), F4/80 (Caltag, San Mateo, CA, USA) antibodies. The fraction occupied by the positively stained areas was calculated from more than 6 high magnification field (HMF) images per group using ImageJ software.
Sanchez-Lopez E., Flashner-Abramson E., Shalapour S., Zhong Z., Taniguchi K., Levitzki A, & Karin M. (2015). Targeting colorectal cancer via its microenvironment by inhibiting IGF-1 Receptor-insulin receptor substrate and STAT3 signaling. Oncogene, 35(20), 2634-2644.
+ Open protocol
+ Expand
10

Assessing STAT3 Phosphorylation in B Cell-Depleted Splenocytes

Check if the same lab product or an alternative is used in the 5 most similar protocols
B cell-depleted splenocytes from normal C57BL/6 mice (4 × 106 cells/ml) were stimulated with/without IL-21 (10 ng/ml, Peprotech, Rocky Hill, NJ) and SM934 (10 μM) for 0 to 60 minutes.
Cell cultures or splenocytes isolated from CIA mice of individual group were directly lysed in sodium dodecyl sulfate (SDS) sample buffer containing protease inhibitor cocktail (Roche Life Science, Mannheim, Germany)57 (link). Whole cell lysates were fractionated on 10% SDS-polyacrylamide gel electrophoresis (PAGE) and Western blotted with antibodies to phosphorylated STAT3 (Cell Signaling Technology, Beverly, MA) and GAPDH (KangChen Biotechnology, China). The densities of the bands were quantified with a computerized densitometer (Image J Launcher, Broken Symmetry Software).
Lin Z.M., Yang X.Q., Zhu F.H., He S.J., Tang W, & Zuo J.P. (2016). Artemisinin analogue SM934 attenuate collagen-induced arthritis by suppressing T follicular helper cells and T helper 17 cells. Scientific Reports, 6, 38115.
+ 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!