Anti-Phospho-STAT3 (Y705) Rabbit Monoclonal Antibody

Western blot analysis of STAT3 using anti-STAT3 antibody (P00007-2).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: mouse NIH/3T3 whole cell lysates,
Lane 3: mouse RAW264.7 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-STAT3 antigen affinity purified monoclonal antibody (Catalog # P00007-2) at 1:5000 overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for STAT3 at approximately 88 kDa. The expected band size for STAT3 is at 88 kDa.

Western blot analysis of Phospho-STAT3 (Y705) using anti-Phospho-STAT3 (Y705) antibody (P00007-2).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: Normal group-rat colon tissue lysates,
Lane 2: Model group-rat colon tissue lysates,
Lane 3: Triditional Chinese medicine treatment (low dose)-rat colon tissue lysates,
Lane 4: Triditional Chinese medicine treatment (medium dose)-rat colon tissue lysates,
Lane 5: Triditional Chinese medicine treatment(high dose)-rat colon tissue lysates,
Lane 6: Western medicine treatment-rat colon tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Phospho-STAT3 (Y705) antigen affinity purified monoclonal antibody (Catalog # P00007-2) at 1:5000 overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a HRP Conjugated AffiniPure Goat Anti-rabbit IgG (H+L) secondary antibody at a dilution of 1:5000 for 1 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with ChemiDoc MP system. A specific band was detected for Phospho-STAT3 (Y705) at approximately 88 kDa. The expected band size for Phospho-STAT3 (Y705) is at 88 kDa.

IHC analysis of STAT3 using anti-STAT3 antibody (P00007-2).
STAT3 was detected in a paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-STAT3 Antibody (P00007-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

Immunofluorescent analysis of HeLa cells treated with IFN-alpha, using Phospho-STAT3 (Y705) Antibody.

Ruxolitinib effectively reverse the PDGFRB Y562D -induced phenotypic modulation in HBVSMCs. Western blotting illustrates the alterations in p-JAK2 and p-STAT (p-STAT1 and p-STAT3) expression across different treatment groups ( A ). After normalization to the control group, the relative density of immunoblot bands in ( A ) is presented in scatter bar graph ( B ). Immunoblots ( C ) and relative density of bands ( D ) of markers associated with phenotypic modulation in HBVSMCs under different treatments are shown. A bar graph is used to present the rt-qPCR results for smooth muscle cell (SMC) markers and inflammatory markers in HBVSMCs under different treatment conditions ( E ). The results of RT-qPCR are analyzed using Tukey's multiple comparisons test. EDU assay demonstrate the proliferative capacity of HBVSMCs receiving different treatments ( F ) and statistical analysis of the proportion of Edu-positive cells in the different groups from 5 different fields of each group at × 200 magnification. Statistical differences are detected using Tukey's multiple comparisons test ( G ). Scratch assay shows the proliferative capacity of HBVSMCs underwent different treatment ( H ) and statistical analysis of the rate of wound healing (reduced area at 4H /area at 0H) in the different groups from 5 different fields of each group at × 200 magnification. Statistical differences are detected using Tukey's multiple comparisons test ( I ). ns, no significant; * p < 0.05; ** p < 0.01. The above experiments are all repeated three times
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GO term, KEGG pathway, and GSEA enrichment analyses of HK2-related genes. A - B Heatmap of the top 50 positively and negatively related genes with HK2. C KEGG pathway enrichment analysis using the GSEA database. D - I Gene set enrichment plots of the NF-κB ( D ), complement and coagulation cascades ( E ), Toll-like receptor ( F ), NOD-like receptor ( G ), Th17 cell differentiation ( H ), and JAK-STAT ( I ) signalling pathways. J - M GO and KEGG enrichment analyses of HK2 positively or negatively related genes (Pearson’s rho ≥0.5, P < 0.05). N Expression levels of p-STAT3, STAT3, p-Akt, Akt in HK2-knockdown and control groups in T98 and GBM1 cell lines . Data shown as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001
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The JAK2-STAT pathway serves as the major downstream signaling pathway of the mutated PDGFB. mIF demonstrate the important downstream signaling pathway markers of PDGFRβ (p-JAK, p-Src, p-Erk1/2 and p-PLCγ) in FIA sections and NCA sections ( A ). Scale bar, 100 μm. Statistical analysis is performed to compare the mean fluorescence intensity of different signal markers between NCAs ( n = 4) and FIAs ( n = 5) ( C ). The expression of p-STAT1 and p-STAT3, which are downstream of p-JAK2, is detected by mIF in FIAs and NCAs. Scale bar, 50 μm ( B ). Statistical analysis of mean fluorescence intensity of these marks between NCAs ( n = 4) and FIAs ( n = 5) ( D ). Western blotting displays the expression of the important downstream signal pathway markers of PDGFRβ (p-JAK2, p-Src, p-Erk1/2 and p-PLCγ) in the HBVSMCs underwent different treatment in vitro ( E ). Relative density (normalized to Control) of immunoblot bands from ( E ) corresponding to pPLCγ, p-JAK2, p-Src, and p-ERK1/2 ( n = 3) ( F ). 'n' represented the number of samples. Three random fields were selected for statistical analysis in each sample, and the average value represented the detection value of this marker in this sample. Tukey's multiple comparisons test is conducted to evaluate the statistical differences. ns, no significant; ***padj < 0.001, ****padj < 0.0001
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Curcumol combined with sorafenib inhibited the tumor growth of Huh7-R cell xenografts in vivo . (A) The gross appearance of the tumors in xenografted mice. (B) The tumor weight of xenografted mice after 14 days of intervention. (C) Tumor volumes in xenograft mice after 7 and 14 days of intervention. (D) Body weight of xenografted mice before and after 14 days of intervention. (E) HE staining of tumors. Scar bar = 100 μm. (F) Ki67 staining of tumors. Scar bar = 100 μm. (G) TUNEL staining of tumors. Scar bar = 50 μm. (H, I) Tumor tissues were stained for p-AKT (H) , and p-STAT3 (I) . Scar bar = 100 μm *p < 0.05, **p < 0.01, ***p < 0.001.
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Experimental verification of the function of Curcumol in Sorafenib-resistant HCC cells. (A) Effect of Sorafenib (5, 10, 15, 20, 30, 40, and 50 μM) on Huh7-R and HepG2-R cells viability. (B) Effect of Curcumol (31.25, 62.5, 125, 250, 500, and 1000 μM) on Huh7-R and HepG2-R cells viability. (C) Effect of Curcumol combined with Sorafenib (5, 10, 15, 20, 30, 40, and 50 μM) on Huh7-R and HepG2-R cells viability. (D) Colony formation assay of Huh7-R and HepG2-R cells treated with Curcumol, Sorafenib, and Curcumol combined with Sorafenib, respectively. (E) Flow cytometry detection of apoptosis analysis of Huh7-R and HepG2-R cells treated with Curcumol, Sorafenib, and Curcumol combined with Sorafenib, respectively. (F) Transwell assay was performed to detect the effect of Curcumol on the migratory ability of Huh7-R cells. (G) Transwell assay to detect the effect of Curcumol on the invasive ability of Huh7-R cells. (H) Cell scratch assay of Huh7-R cells treated with Curcumol, Sorafenib, and Curcumol combined with Sorafenib, respectively. (I) The protein expression levels of PI3K\AKT pathway and JAK/STAT3 pathway following intervention with Curcumol combined with Sorafenib. *p < 0.05, **p < 0.01, ***p < 0.001.
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Relationship between differentially expressed core targets and immune cell infiltration. (A) ALB (B) STAT3 (C) HSP90AA1 (D) HSP90AB1 (E) SRC.
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Molecular docking pattern of Curcumol with target proteins. (A) ALB (B) STAT3 (C) HSP90AA1 (D) HSP90AB1 (E) SRC.
Index in PubMed under a CC BY license. PMID: 40242448