Cells seeded into a 6-well plate were treated with 30 ng/ml human IL-6 or not for the indicated time. The cytoplasm and nuclear fraction of cells were extracted using a nuclear and cytoplasmic protein extraction kit (Beyotime, Shanghai, China). Whole cell lysates or the nuclear/cytoplasm fractions were subjected to SDS-PAGE and immunoblotting [14 (link)]. Primary antibodies against STAT3 (Abcam, USA), phosphorylated STAT3 (p-STAT3) (Abcam, USA), β-actin (Abcam, USA), GAPDH (a cytoplasm fraction maker, Abcam, USA), Histone H3 (a nuclear fraction maker, Abcam, USA), and flag (Beyotime, China) were used.
Phosphorylated stat3 p stat3
Manufactured by Abcam
Sourced in United Kingdom
Phosphorylated STAT3 (p-STAT3) is a protein that has been modified by the addition of a phosphate group. STAT3 (Signal Transducer and Activator of Transcription 3) is a transcription factor that plays a role in cellular processes such as cell growth, differentiation, and survival. Phosphorylation of STAT3 is an important regulatory mechanism that can influence its activity and function.
Automatically generated - may contain errors
Lab products found in correlation
Gapdh, by Abcam (2 mentions)
β actin, by Abcam (2 mentions)
Histone h3, by Abcam (1 mentions)
Nuclear and cytoplasmic protein extraction kit, by Beyotime (1 mentions)
Flag tag (flag), by Beyotime (1 mentions)
Dimethyl sulphoxide, by Merck Group (1 mentions)
Vimentin, by Abcam (1 mentions)
Twist1, by Abcam (1 mentions)
Oridonin c20h28o6, by Merck Group (1 mentions)
Phosphorylated akt p akt, by Abcam (1 mentions)
Horseradish peroxidase conjugated goat anti rabbit secondary antibody, by Cell Signaling Technology (1 mentions)
E cadherin, by Abcam (1 mentions)
Iba 1, by Abcam (1 mentions)
Stat3, by Abcam (1 mentions)
Nlrp3, by Cell Signaling Technology (1 mentions)
Cd11b, by Abcam (1 mentions)
Caspase 1, by Cell Signaling Technology (1 mentions)
3 protocols using phosphorylated stat3 p stat3
1
STAT3 Activation under IL-6 Treatment
2
Oridonin Inhibits Cancer Cell Migration
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Oridonin (C20H28O6), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and dimethyl sulphoxide (DMSO) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Antibodies against AKT, phosphorylated-AKT (p-AKT), STAT3, phosphorylated-STAT3 (p-STAT3), E-cadherin, Vimentin, Twist1 and β-actin were obtained from Abcam (Cambridge, UK). Phalloidin dye solution, horseradish peroxidase-conjugated goat anti-rabbit secondary antibody were purchased from Cell Signaling Technology, Inc (Beverly, MA, USA).
3
Protein Expression Analysis in Cells
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After a 20-minute centrifugation at 12,000 g, the protein concentrations of the supernatant were determined. The samples (30 μg per sample) were loaded into a 10-12% sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Next, after transferring onto a polyvinylidene difluoride (PVDF) membrane, the protein bands were blocked in 5% fat-free milk for 1 h at RT. Then, after overnight incubation with the indicated primary antibodies at 4°C, washing 3 times with phosphate-buffered saline with Tween® detergent (PBST), and a 1-hour incubation with horseradish peroxidase (HRP)-conjugated secondary antibodies at RT, the protein bands and signals were detected using an enhanced chemiluminescence detection system (Pierce, Rockford, USA). The antibodies used were as follows: GAPDH (1:1000; Abcam, Cambridge, UK), phosphorylated STAT3 (p-STAT3, 1:1000; Abcam), STAT3 (1:1000; Abcam), ASC (1:1000; Cell Signaling Technology, Danvers, USA), caspase-1 (1:800; Cell Signaling Technology), NLRP3 (1:1000; Cell Signaling Technology), ionized calcium-binding adapter molecule 1 (Iba-1, 1:800; Abcam), and cluster of differentiation molecule 11b (CD11b, 1:1000; Abcam). All protein expressions were normalized to GAPDH.
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