Log In Sign up
The largest database of trusted experimental protocols
> Chemicals & Drugs > Amino Acid > STAT3 protein, human

STAT3 protein, human

STAT3 (Signal Transducer and Activator of Transcription 3) is a member of the STAT family of transcription factors.
It plays a crucial role in cellular processes such as cell growth, differentiation, and apoptosis.
STAT3 is activated by phosphorylation and translocates to the nucleus, where it binds to specific DNA response elements and regulates the expression of target genes.
Dysregulation of STAT3 has been implicated in various diseases, including cancer, inflammation, and autoimmunity.
Researchers studying STAT3 protein can leverage the power of AI-driven protocol optimization to enhance the reproducibility and efficacy of their experiments, utilizing the best protocols from literature, preprints, and patents identified through PubCompare.ai's advanced comparisons.

Most cited protocols related to «STAT3 protein, human»

1
Cloning and Transfection of STAT3 and HMGA1a Vectors
Cited 16 times
The pSG5-STAT3-C vector was made by excision of constitutively activated STAT3 (STAT3-C) from STAT3 pcDNA3.1 vector (14 (link)) with Pme1 and BamH1. The STAT3-C fragment was cloned into pSG5 after restriction with EcoR1 and Klenow treatment. The PA-HMGA1a vector was made by excision of the full-length murine HMGA1a cDNA from the vector pHEBoNeo-HMG-I3 with HindIII, Klenow treatment, and subsequent NotI digestion. The HMGA1a fragment was subsequently cloned into the EF1α expression vector (18 (link)) after restriction with BamHI and Klenow treatment. The STAT promoter vectors were described (kindly provided by K. Kohno, Department of Molecular Biology, School of Medicine, University of Occupational and Environmental Health, Kitakyushu, Fukuoka, Japan; ref. 19 (link)).
Transfections were done using Lipofectamine 2000 (Invitrogen) according to the manufacturer’s instructions as we previously described (10 (link)). In the experiments with HEL control or HEL-HMGA1 cells, the STAT3 promoter reporter plasmid (1 μg) was cotransfected with the control pRL-TK vector (100 ng) containing Renilla luciferase (Promega) to control for transfection efficiency (3 × 105 cells/500 μL per transfection). The DNA was mixed with Lipofectamine 2000 at a ratio of 1 μg:1.5 μL (in Opti-MEM, Invitrogen) and incubated with cells for 6 h. Cells were harvested for luciferase activity 24 h after transfection and experiments were done twice in triplicate. In the titration experiment, reporter plasmid (100 ng), control Renilla luciferase plasmid (50 ng), and 0 to 150 ng of HMGA1a expression plasmid (pSG5-HMG-I) were cotransfected as described above. The pSG5 plasmid was added (0–150 ng) to keep the total DNA constant in the titration experiments (1 × 105 cells/100 μL per transfection). The DNA was mixed with Lipofectamine 2000 at a ratio of 1 μg:1.67 μL (in Opti-MEM, Invitrogen) and cells were harvested as described above.
The lentiviral construct expressing dominant-negative STAT3 (Lenti-DN-STAT3) was made from the pcDNA3.1-STAT3DN (provided by J-I. Park, the Johns Hopkins University School of Medicine, Baltimore, MD; ref. 14 (link)). The cytomegalovirus promoter was replaced by the human EF1α promoter from pEF1/Myc-His-A (Invitrogen) and the EF-DN-STAT3 cassette was cloned into the cFUGW lentivirus (provided by D. Baltimore, Division of Biology, California Institute of Technology, Pasadena, CA) at the PacI site and lentiviral supernatants were made as described (20 (link)).
Hillion J., Dhara S., Sumter T.F., Mukherjee M., Di Cello F., Belton A., Turkson J., Jaganathan S., Cheng L., Ye Z., Jove R., Aplan P., Lin Y.W., Wertzler K., Reeves R., Elbahlouh O., Kowalski J., Bhattacharya R, & Resar L.M. (2008). The High-Mobility Group A1a/Signal Transducer and Activator of Transcription-3 Axis: An Achilles Heel for Hematopoietic Malignancies?. Cancer research, 68(24), 10121-10127.
Publication 2008
Cells Cloning Vectors Cytomegalovirus Digestion DNA, Complementary HMGA1 protein, human Homo sapiens Lentivirus lipofectamine 2000 Luciferases Luciferases, Renilla Mus Pharmaceutical Preparations Plasmids Promega STAT3 protein, human Titrimetry Transfection
2
Validating STAT3 Mutations by Sanger
Cited 13 times
After exome sequencing, the candidate mutations were validated with the use of Sanger sequencing. For the STAT3 mutation screening, five primer pairs were designed to cover the six exons encoding the Src homology 2 (SH2) domain of STAT3 (exons 19 to 24). (The primers are described in Table S3 in the Supplementary Appendix.)
Koskela H.L., Eldfors S., Ellonen P., van Adrichem A.J., Kuusanmäki H., Andersson E.I., Lagström S., Clemente M.J., Olson T., Jalkanen S.E., Majumder M.M., Almusa H., Edgren H., Lepistö M., Mattila P., Guinta K., Koistinen P., Kuittinen T., Penttinen K., Parsons A., Knowles J., Saarela J., Wennerberg K., Kallioniemi O., Porkka K., Loughran TP J.r., Heckman C.A., Maciejewski J.P, & Mustjoki S. (2012). Somatic STAT3 Mutations in Large Granular Lymphocytic Leukemia. The New England journal of medicine, 366(20), 1905-1913.
Publication 2012
Exons Mutation Oligonucleotide Primers src Homology Domains STAT3 protein, human
3
Evaluating STAT3 Phosphorylation Dynamics
Cited 12 times
Spectral characteristics of AEC, DAB and NovaRed were displayed with the different color models using color bars that were generated from a range of representative IHC staining derived from the xenograft studies described below, as well as additional IHC archived slides that showed a greater range of staining intensity.
Changes in STAT3 phosphorylation (p-STAT3) levels were associated with different conditions, including growth factor stimulation and sampling techniques as previously described [16 ]. Formalin-fixed paraffin embedded specimens were obtained to compare IHC staining using different chromogens AEC, DAB or NovaRed, and hematoxylin counterstain. Briefly, a tumor xenograft was excised and divided into two equal parts. One part was further divided and subjected to immediate formalin fixation, or protein extraction for Western blot analysis [17 (link)]. The second part was incubated with 50 ng/ml epidermal growth factor (EGF, Sigma-Aldrich, Oakville, ON) in PBS for 20 minutes prior to specimen analysis. The antibody S727p-STAT3 (BD Biosciences, Mississauga, ON) was used for Western blotting at a dilution of 1:100, and anti-GAPDH used at a dilution of 1:5000 (Ambion, Foster City, CA). Protein bands were detected with ECL plus fluorescence, imaged with a Typhoon system and their intensities were quantified (mean intensity × band area) and normalized with GAPDH using ImageQuant5.2 software (GE Healthcare, Little Chalfont, Buckinghamshire, UK).
Second, p-STAT3 levels were also quantified in formalin-fixed paraffin embedded sections of fine-needle samples obtained from a tumor xenograft. These samples were subjected to either immediate or delayed fixation after 5, 20 or 60 minutes, which caused an increase in p-STAT3.
Pham N.A., Morrison A., Schwock J., Aviel-Ronen S., Iakovlev V., Tsao M.S., Ho J, & Hedley D.W. (2007). Quantitative image analysis of immunohistochemical stains using a CMYK color model. Diagnostic Pathology, 2, 8.
Full text: Click here
Publication 2007
ADRB2 protein, human associated conditions azo rubin S Epidermal growth factor Fluorescence Formalin GAPDH protein, human Growth Factor Hematoxylin Heterografts Immunoglobulins Needles Neoplasms Paraffin Paraffin Embedding Phosphorylation Proteins STAT3 protein, human Technique, Dilution Typhoons Western Blot
4
Leptin Signaling Pathway in Obesity
Cited 11 times
A detailed Methods section is available in the online supplement at http://circres.ahajournals.org.
Adenovirus expressing an enhanced green fluorescence protein (Ad-GFP), used as control, or both Ad-Cre and Ad-GFP (referred to as Ad-Cre) were microinjected bilaterally into the ARC of ObRflox/flox mice. Leptin activation of Stat3 was analyzed by immunohistochemistry. Recording of SNA was obtained in anesthetized mice. Obesity was induced by high fat (45% kcal) feeding for 10 weeks.
Harlan S.M., Morgan D.A., Agassandian K., Guo D.F., Cassell M.D., Sigmund C.D., Mark A.L, & Rahmouni K. (2011). Ablation of the Leptin Receptor in the Hypothalamic Arcuate Nucleus Abrogates Leptin-Induced Sympathetic Activation. Circulation research, 108(7), 808-812.
Publication 2011
Adenovirus Vaccine Dietary Supplements enhanced green fluorescent protein Immunohistochemistry Leptin Mice, Laboratory Obesity STAT3 protein, human
5
Identifying Immumomodulatory Drug Targets
Cited 9 times
We inspected ENCODE data on UCSC genome browser to identify putative IL2 enhancers in one or more cell types that may be responsive to FDA approved immomodulatory drugs. We scanned the intergenic region upstream of IL2 in hg19 for (i) enhancer-associated histone marks (H3K4me1 and H3K27ac), (ii) binding by one or more TFs as confirmed by ChIP-seq, and (iii) the TF pathway can be targeted by a human therapeutic. This analysis identified IRF4 and STAT3 binding sites in addition to the known NFAT-responsive elements (main text ref 28 (link)).
Buenrostro J.D., Giresi P.G., Zaba L.C., Chang H.Y, & Greenleaf W.J. (2013). Transposition of native chromatin for multimodal regulatory analysis and personal epigenomics. Nature methods, 10(12), 1213-1218.
Publication 2013
Binding Sites Cells Chromatin Immunoprecipitation Sequencing Genome Histone Code Homo sapiens interferon regulatory factor 4, human Intergenic Region Pharmaceutical Preparations STAT3 protein, human Therapeutics

Most recents protocols related to «STAT3 protein, human»

1
Preparing ERK1/2 and STAT3 Inhibitors
ERK1/2 inhibitor (U0126 solution): U0126 powder was dissolved in dimethyl sulfoxide (DMSO) to 50µM concentration as the storage solution.
STAT3 inhibitor (Stattic solution): Stattic powder was dissolved in DMSO to 20µM concentration as the storage solution.
Xiao X., Zou S, & Chen J. (2023). Cyclic tensile force modifies calvarial osteoblast function via the interplay between ERK1/2 and STAT3. BMC Molecular and Cell Biology, 24, 9.
Full text: Click here
Publication 2023
Mitogen-Activated Protein Kinase 3 Powder STAT3 protein, human stattic Sulfoxide, Dimethyl U 0126
2
Co-immunoprecipitation of ERK1/2 and STAT3
The total protein was extracted as mentioned before. Co-IP was conducted as described previously [27 (link)]. 10% of the supernatants were prepared for Input, and the rest were incubated with normal IgG, anti-ERK1/2 (dilution 1:20), or anti-STAT3 (dilution 1:20) at 4 °C overnight with gentle end-over-end mixing, followed by pre-washed Protein A/G magnetic beads (Shanghai Epizyme Biomedical Technology Co., Ltd), mellow agitation, and overnight attachment. Protein A/G magnetic beads conjugates were separated by a magnetic rack. Proteins were eluted by boiling with loading buffer and then detached from beads by the magnetic rack.
Xiao X., Zou S, & Chen J. (2023). Cyclic tensile force modifies calvarial osteoblast function via the interplay between ERK1/2 and STAT3. BMC Molecular and Cell Biology, 24, 9.
Full text: Click here
Publication 2023
Biomedical Technology Buffers G-substrate Mitogen-Activated Protein Kinase 3 Proteins Staphylococcal Protein A STAT3 protein, human Technique, Dilution
3
Protein Expression Analysis in Osteogenic Differentiation

Total cellular protein (Signalway Antibody LLC, USA) and nuclear protein (Shanghai Sangon Biotechnology Co., Ltd, China) were extracted using the corresponding kit. The sample concentration was determined by a BCA protein assay kit (Beyotime, China). After being mixed with loading buffer, the protein sample was boiled at 95 °C for 5 min. The protein sample was separated by SDS polyacrylamide gel electrophoresis and transferred to PVDF membrane. Later, the blots were blocked in 5% fat-free skimmed milk for one hour at room temperature, followed by incubation with primary antibodies (Huabio, China) against Osterix (1:1000, ER1914-47), COL-I (1:1000, ET1609-68), OCN (1:1000, ER1919-20), Runx2 (1:1000, ET1612-47), ALP (1:1000, ET1601-21), ERK1/2(1:5000, ET1601-29), pERK1/2 (1:1000, ET1603-22), STAT3(1:1000, ET1607-38), pSTAT3(1:1000, ET1603-40), and GAPDH (1:1000, ET1601-4 ) overnight at 4 °C. After a three-time wash in TBST solution, membranes were probed with goat anti-rabbit IgG conjugated secondary antibody (Hubio, China) for one hour at room temperature. The blots were cut prior to hybridization with antibodies during blotting. The membranes were processed with enhanced chemiluminescence detection reagents (Applygen Technologies Inc., Beijing, China) and visualized.
Xiao X., Zou S, & Chen J. (2023). Cyclic tensile force modifies calvarial osteoblast function via the interplay between ERK1/2 and STAT3. BMC Molecular and Cell Biology, 24, 9.
Full text: Click here
Publication 2023
anti-IgG Antibodies Biological Assay Buffers Cardiac Arrest Cells Chemiluminescence Crossbreeding GAPDH protein, human Goat Immunoglobulins Milk, Cow's Mitogen-Activated Protein Kinase 3 Nuclear Protein polyvinylidene fluoride Proteins Rabbits RUNX2 protein, human SDS-PAGE Staphylococcal Protein A STAT3 protein, human Tissue, Membrane
4
Western Blot Analysis of Apoptotic Signaling
MDCK cells were grown
to 80% on 10 cm dishes, incubated with H3N2 for 2 h, and then treated
with 16 μM Se@CS for 48 h. Total protein was extracted, and
the protein concentration was determined using the BCA kit. After
denaturation at 95 °C for 10 min, the protein samples were separated
by 10% SDS-PAGE gel and transferred to a difluoride film in polyvinylidene.
Polyvinylidene difluoride membranes were sealed with 10% BSA for 2
h and incubated overnight with primary antibodies including JNK, p53,
caspase-3, BAD, JAK, p-STAT3, Akt, and β-actin antibodies. Then,
the membrane was washed with TBST and incubated with the secondary
antibody for 2 h. After the membrane was fully mixed with luminescent
solution, the protein band images were exposed and preserved with
a gel imager, and the exposure process should be carried out without
light.
Xu T., Lai J., Su J., Chen D., Zhao M., Li Y, & Zhu B. (2023). Inhibition of H3N2 Influenza Virus Induced Apoptosis by Selenium Nanoparticles with Chitosan through ROS-Mediated Signaling Pathways. ACS Omega, 8(9), 8473-8480.
Publication 2023
Actins Antibodies CASP3 protein, human Hyperostosis, Diffuse Idiopathic Skeletal Madin Darby Canine Kidney Cells polyvinylidene fluoride Proteins SDS-PAGE STAT3 protein, human Tissue, Membrane
5
Liver Tissue Western Blot Analysis
The liver tissues were homogenized and lysed in ice-cold lysis buffer supplemented with protease and phosphatase inhibitors (Topscience, Shanghai, China) and centrifuged at 12,000 g for 20 min at 4°C, and then the clarified supernatants were collected. Then proteins were quantified with the Pierce BCA Protein Assay Kit (Thermo, United States) assay according to the manufacturer’s instructions. The western blot was performed as previously described (Li et al., 2022a (link); Li et al., 2022b (link)). Briefly, after SDS-PAGE and transmembrane, the target proteins were accordingly probed with first antibodies against GAPDH (10494-1-AP, 1:1,000, Proteintech), phospho-Stat3 (9145, 1:1,000, CST), Stat3 (9139, 1:1,000, CST), phospho-p38 MAPK (4511, 1:1,000, CST), p38 MAPK (8690, 1:1,000, CST), phospho-NF-κB p65 (3033, 1:1,000, CST), NF-κB p65 (6956, 1:1,000, CST), phospho-IκBα (2859, 1:1,000, CST), and IκBα (4814, 1:1,000, CST), respectively. After incubation with the corresponding HRP-conjugated secondary antibody, the signal of the target protein was detected using a ChemiDo XRS gel imager system (Bio-Rad, United States) with Immobilon Western Chemiluminescent HRP Substrate (Millipore, United States) and was scanned by ImageJ software. The ratio of the target protein was normalized to the internal control protein GAPDH, and fold-change was calculated relative to the control group.
Sun H., Wang X.K., Li J.R., Tang M., Li H., Lei L., Li H.Y., Jiang J., Li J.Y., Dong B., Jiang J.D, & Peng Z.G. (2023). Establishment and application of a high-throughput screening model for cell adhesion inhibitors. Frontiers in Pharmacology, 14, 1140163.
Full text: Click here
Publication 2023
alpha, NF-KappaB Inhibitor Antibodies Biological Assay Buffers Cold Temperature Endopeptidases GAPDH protein, human Immobilon Immunoglobulins inhibitors Liver Mitogen-Activated Protein Kinase p38 Phosphoric Monoester Hydrolases Proteins Protein Targeting, Cellular SDS-PAGE STAT3 protein, human Tissues Transcription Factor RelA Western Blotting

Top products related to «STAT3 protein, human»

1
Stat3 by Cell Signaling Technology
Sourced in United States, United Kingdom, China, Germany, Japan, Morocco
STAT3 is a protein that plays a key role in cell signaling pathways. It functions as a transcription factor, regulating the expression of genes involved in various cellular processes such as cell growth, differentiation, and survival.
2
Lipofectamine 2000 by Thermo Fisher Scientific
Sourced in United States, China, Germany, United Kingdom, Canada, Japan, France, Italy, Switzerland, Australia, Spain, Belgium, Denmark, Singapore, India, Netherlands, Sweden, New Zealand, Portugal, Poland, Israel, Lithuania, Hong Kong, Argentina, Ireland, Austria, Czechia, Cameroon, Taiwan, Province of China, Morocco
Lipofectamine 2000 is a cationic lipid-based transfection reagent designed for efficient and reliable delivery of nucleic acids, such as plasmid DNA and small interfering RNA (siRNA), into a wide range of eukaryotic cell types. It facilitates the formation of complexes between the nucleic acid and the lipid components, which can then be introduced into cells to enable gene expression or gene silencing studies.
3
P stat3 by Cell Signaling Technology
Sourced in United States, United Kingdom, China, Germany, Macao
P-STAT3 is an antibody that specifically detects phosphorylated STAT3 protein. STAT3 (Signal Transducer and Activator of Transcription 3) is a transcription factor that plays a key role in cellular signaling pathways. Phosphorylation of STAT3 is a crucial step in its activation and regulation of target gene expression.
4
Pvdf membrane by Merck Group
Sourced in United States, Germany, China, United Kingdom, Morocco, Ireland, France, Italy, Japan, Canada, Spain, Switzerland, New Zealand, India, Hong Kong, Sao Tome and Principe, Sweden, Netherlands, Australia, Belgium, Austria
PVDF membranes are a type of laboratory equipment used for a variety of applications. They are made from polyvinylidene fluoride (PVDF), a durable and chemically resistant material. PVDF membranes are known for their high mechanical strength, thermal stability, and resistance to a wide range of chemicals. They are commonly used in various filtration, separation, and analysis processes in scientific and research settings.
5
Anti stat3 by Cell Signaling Technology
Sourced in United States, United Kingdom, Germany, China
Anti-STAT3 is a primary antibody product from Cell Signaling Technology. It is designed to detect STAT3 (signal transducer and activator of transcription 3), a transcription factor involved in various cellular processes. This antibody can be used for applications such as Western blotting, immunoprecipitation, and immunohistochemistry.
6
Fetal bovine serum (fbs) by Thermo Fisher Scientific
Sourced in United States, China, United Kingdom, Germany, Australia, Japan, Canada, Italy, France, Switzerland, New Zealand, Brazil, Belgium, India, Spain, Israel, Austria, Poland, Ireland, Sweden, Macao, Netherlands, Denmark, Cameroon, Singapore, Portugal, Argentina, Holy See (Vatican City State), Morocco, Uruguay, Mexico, Thailand, Sao Tome and Principe, Hungary, Panama, Hong Kong, Norway, United Arab Emirates, Czechia, Russian Federation, Chile, Moldova, Republic of, Gabon, Palestine, State of, Saudi Arabia, Senegal
Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.
7
Gapdh by Cell Signaling Technology
Sourced in United States, United Kingdom, China, Germany, Japan, Canada, Morocco, France, Netherlands, Ireland, Israel, Australia, Sweden, Macao, Switzerland
GAPDH is a protein that functions as an enzyme involved in the glycolysis process, catalyzing the conversion of glyceraldehyde 3-phosphate to 1,3-bisphosphoglycerate. It is a common reference or housekeeping protein used in various assays and analyses.
8
Ab76315 by Abcam
Sourced in United States, United Kingdom, China
Ab76315 is a laboratory equipment product offered by Abcam. It is a device designed for specific laboratory functions. The core function of this product is to facilitate laboratory procedures, but a detailed description cannot be provided while maintaining an unbiased and factual approach.
9
Phospho stat3 by Cell Signaling Technology
Sourced in United States, United Kingdom, China, Morocco, Germany
Phospho-STAT3 is a laboratory reagent that detects the phosphorylation of Signal Transducer and Activator of Transcription 3 (STAT3), a transcription factor involved in various cellular processes. This product allows researchers to study the activation and regulation of STAT3 signaling pathways.
10
Stat3 by Abcam
Sourced in United States, United Kingdom, China
STAT3 is a transcription factor that mediates the expression of genes involved in cell growth and survival. It is a member of the STAT (Signal Transducer and Activator of Transcription) family of proteins and plays a crucial role in various cellular processes, including immune response, cell differentiation, and cancer development.

More about "STAT3 protein, human"

STAT3 (Signal Transducer and Activator of Transcription 3) is a crucial player in cellular processes like growth, differentiation, and apoptosis.
As a member of the STAT family of transcription factors, STAT3 gets activated through phosphorylation and then translocates to the nucleus.
There, it binds to specific DNA response elements, regulating the expression of target genes.
Dysregulation of STAT3 has been implicated in various diseases, including cancer, inflammation, and autoimmunity.
To enhance the reproducibility and efficacy of STAT3 protein research, scientists can leverage the power of AI-driven protocol optimization.
PubCompare.ai's advanced comparisons can help researchers identify the best protocols from literature, preprints, and patents.
This allows them to utilize the optimal products and procedures, elevating their experiments and boosting the quality of their findings.
Some key tools and techniques commonly used in STAT3 protein research include Lipofectamine 2000 for transfection, P-STAT3 antibodies for detecting phosphorylated STAT3, PVDF membranes for Western blotting, and Anti-STAT3 antibodies for immunoprecipitation.
Additionally, researchers may use FBS (Fetal Bovine Serum) as a cell culture supplement, and employ GAPDH as a loading control.
The Ab76315 antibody is a commonly used tool for STAT3 detection.
By optimizing their protocols and leveraging the best available resources, scientists can enhance the reproducibility and impact of their STAT3 protein research, leading to breakthroughs in understanding cellular processes and developing targeted therapies for related diseases.