Reagents were purchased as follows: 5% imiquimod cream manufactured by Perrigo (Yeruham, Israel), calcipotriol (MC903) from Cayman Chemical (Ann Arbor, MI), sirius red (Direct Red 80) from Sigma-Aldrich (St. Louis, MO), anti-phospho-RelA(S536), anti-TNFAIP3 (A20), anti-GAPDH, anti-phospho-Stat3 (Y705), and anti-phospho-Stat6 (Y641) from Cell Signaling (Danvers, MA), anti-filaggrin antibody from Santa Cruz Biotech (Dallas, TX), anti-keratin 5 antibody from LifeSpan Biosciences (Seattle, WA), anti-GATA3 and anti-IL23R from Abcam (Cambridge, MA), anti-CD4 from eBioscience (San Diego, CA), and NovaUltra™ Toluidine Blue Stain from Fisher Scientific (Pittsburgh, PA). Rabbit anti-Trim32 antibody for immunoblotting and chicken anti-Trim32 antibody for immunostaining were generated in our laboratory (Albor et al. 2006 (link)).
Anti phospho stat3 y705
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-phospho-STAT3 (Y705) is a primary antibody that specifically recognizes the phosphorylated form of STAT3 at tyrosine 705. STAT3 is a transcription factor that plays a critical role in cellular signaling pathways.
Automatically generated - may contain errors
Lab products found in correlation
Anti stat3, by Cell Signaling Technology (15 mentions)
Anti gapdh, by Santa Cruz Biotechnology (5 mentions)
Anti stat1, by Cell Signaling Technology (4 mentions)
Anti phospho stat1 y701, by Cell Signaling Technology (4 mentions)
Anti β actin, by Merck Group (4 mentions)
Anti cleaved caspase 3, by Cell Signaling Technology (3 mentions)
Anti β actin, by Cell Signaling Technology (3 mentions)
Anti caspase 3, by Cell Signaling Technology (3 mentions)
Anti sox2, by Cell Signaling Technology (3 mentions)
Anti β actin, by Santa Cruz Biotechnology (3 mentions)
Anti phospho akt s473, by Cell Signaling Technology (3 mentions)
Anti stat5, by Santa Cruz Biotechnology (2 mentions)
Anti c myc tag, by Fortis Life Sciences (2 mentions)
Horseradish peroxidase conjugated anti rabbit igg, by Thermo Fisher Scientific (2 mentions)
Anti stat3, by Santa Cruz Biotechnology (2 mentions)
Anti iκbα, by Cell Signaling Technology (2 mentions)
Anti cdk2, by Cell Signaling Technology (2 mentions)
Anti cdk4, by Cell Signaling Technology (2 mentions)
Anti cyclin d1, by Cell Signaling Technology (2 mentions)
Anti bax, by Cell Signaling Technology (2 mentions)
29 protocols using anti phospho stat3 y705
1
Inflammatory Skin Disease Protocol
2
Signaling Pathway Analysis by Western Blotting
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The cell lysates were subjected to SDS-PAGE, then the signaling properties were analyzed by western blotting. The list of rabbit primary antibodies used in this study is described as follows: anti phospho-STAT1 (Y701)(Cell Signaling Technology, Danvers, MA), anti STAT1 (Cell Signaling Technology), anti phospho-STAT3 (Y705) (Cell Signaling Technology), anti STAT3 (Santa Cruz Biotechnology, Santa Cruz, CA), anti phospho-STAT5 (Y694) (Cell Signaling Technology), anti STAT5 (Santa Cruz Biotechnology), anti HA tag (BETHYL), anti V5 tag (Millipore, Burling, MA), anti c-myc tag (BETHYL, Montgomery, TX), and anti GAPDH (Santa Cruz Biotechnology). As a secondary antibody, horseradish peroxidase-conjugated anti rabbit IgG (Thermo Fisher Scientific) was commonly used.
3
Protein Expression Profiling by Western Blot
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Protein extracts were loaded onto and separated by SDS-PAGE resolution (Pulilai Co., Beijing, China), followed by transferred onto a PVDF membrane. Next, membrane was blocked and incubated with primary antibodies and HRP-conjugated secondary antibody. Primary antibodies used in current experiments were listed: anti-NF-κB (Abcam, ab32536), anti-phospho-NF-κB (S536) (Abcam, ab76302), anti-IκBα (Cell Signaling Technology, 4812), anti-phospho-IκBα (S32) (Cell Signaling Technology, 2859), anti-TRAF2 (Cell Signaling Technology, 4712), anti-STAT3 (Cell Signaling Technology, 12640), anti-phospho-STAT3 (Y705) (Cell Signaling Technology, 9145), anti-IFNGR2 (ABclonal Technology, A7558), and anti-PD-L1 (Proteintech, 66248-1-Ig). β-actin was applied as a control for protein loading. Gels were visualized using the ECL system (Amersham Biosciences, Uppsala, Sweden) to determine the expression level of targeted proteins.
4
Activation of IL-27 Signaling Pathway
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Lipopolysaccharide (LPS; from Escherichia coli 0127:B8) was purchased from Sigma-Aldrich. Recombinant human cytokine IL-27 was purchased from R&D Systems. Antibody against IL-27RA (Novus Cat# NBP1-02708, RRID:AB_2125066) was purchased from Novus. Anti-phospho-STAT1(Y701) (Cell Signaling Technology Cat# 9171, RRID:AB_331591), anti-phospho-STAT3(Y705) (Cell Signaling Technology Cat# 4113, RRID:AB_2198588), anti-STAT1 (Cell Signaling Technology Cat# 9172, RRID:AB_2198300), and anti-STAT3 (Cell Signaling Technology Cat# 9132, RRID:AB_331588) antibodies were purchased from Cell Signaling Technology. Anti-TLR4 (Abcam Cat# ab13556, RRID:AB_300457), anti-IL-6 (Abcam Cat# 1457-1, RRID:AB_562150), and anti-IL-8 (Abcam Cat# ab34100, RRID:AB_775629) antibodies were purchased from Abcam. Beta(β)-actin (Millipore Cat# MABT825, RRID:AB_2571580) and horseradish peroxidase linked anti-rabbit (Millipore Cat# AP307P, RRID:AB_92641), anti-mouse (Millipore Cat# AP308P, RRID:AB_92635) and anti-goat (Millipore Cat# AP106P, RRID:AB_92411) secondary antibodies were purchased from Millipore. Secondary antibodies for IHC were from Dako.
5
Western Blot Analysis of Cellular Proteins
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Total cell lysates were prepared in 2× sample loading buffer (250 mM Tris-HCl (pH 6.8), 10% glycerol, 4% sodium dodecyl sulfate (SDS), 2% β-mercaptoethanol, 0.006% bromophenol blue, 5 mM sodium orthovanadate, and 50 mM sodium fluoride; Bio-Rad, Hercules, CA, USA). Protein concentrations were quantified through the BCA method [24 (link)] using the BCA Protein Assay Kit (Thermo Fisher Scientific, Waltham, MA, USA). Equal amounts of protein (5–20 µg) were separated using 6–13% SDS-polyacrylamide gel electrophoresis (PAGE) and transferred to polyvinylidene fluoride membranes (Millipore, Bedford, MA, USA). The membranes were blocked with 5% BSA (Sigma-Aldrich) and probed with anti-phospho-STAT3 (Y705), anti-STAT3, anti-cyclin E, anti-CDK2, anti-cyclin D1, anti-CDK4, anti-CDK6, anti-survivin, anti-cleaved caspase-9, anti-cleaved caspase-3, anti-cleaved PARP (D214), and anti-vimentin antibodies (Cell Signaling Technology, Beverly, MA, USA); anti-β-actin horseradish peroxidase antibody (Santa Cruz Biotechnology, Dallas, TX, USA); anti-E-cadherin and anti-N-cadherin antibodies (BD Biosciences, San Jose, CA, USA); or anti-Ki-67 antibody (Abcam, Cambridge, UK). The blots were detected using the WEST-Queen detection system (iNtRON Biotechnology, Seongnam, Korea) [25 (link)].
6
Molecular Mechanisms of Honokiol in Cancer
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Honokiol (≥ 98% of purity) was purchased from LKT laboratories (Minneapolis, MN). Anti-phospho-EGFR (Y1068), anti-total EGFR, anti-phospho-Akt (S473), anti-total Akt, anti-phospho-ERK (T202/Y204), anti-total ERK, anti-phospho-STAT3 (Y705), anti-total STAT3, anti-cyclin D1, anti-CDK2, anti-CDK4, anti- phospho-Rb (S807/811), anti-p27, anti-caspase 3, anti-phospho-GSK3α/β (S21/9), anti-IκBα, anti-bax, anti-phospho-Bad, anti-β-actin, anti-caspase 8, anti-caspase 9 and goat anti-rabbit IgG secondary antibody were from Cell Signaling Technology (Beverly, MA). Anti-poly (ADP-ribose) polymerase (PARP) and anti-p21 were obtained from Santa Cruz Biotechnology. Erlotinib (99% of purity) was purchased from LC Laboratories (Woburn, MA). NNK (99% of purity) was synthesized as described elsewhere [60 (link)].
7
Immunoblot Analysis of STAT3 in GSCs
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Protein from GSCs alone or after 48 h in coculture with astrocytes was collected and subjected to immunoblot analyses as described previously 22 (link). Primary antibodies included anti-STAT3, anti-phospho-STAT3 (Y705) (Cell Signaling Technology, Danvers, MA), and anti-β-actin (Sigma-Aldrich). Donkey anti-rabbit and sheep anti-mouse horseradish peroxidase–conjugated secondary antibodies were purchased from GE Healthcare Life Sciences, Pittsburgh, PA.
8
Protein Expression Quantification Protocol
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Anti-phospho-S6 (S235/236, 1:500), anti-S6 (1:1000), anti-phospho-4E-BP (T37/46, 1:500), anti-4E-BP (1:1000), anti-phospho-STAT3 (Y705, 1:500) and anti-STAT3 (1:500) antibodies were purchased from Cell Signaling Technology (Danvers, MA). Anti-Actin (1:10,000) antibodies, phosphatase Inhibitor Cocktails 2 and 3, and dimethyl sulfoxide (DMSO) were purchased from Sigma Aldrich (St. Louis, MO). The nitrocellulose membrane was purchased from EMD Millipore (Billerica, MA, USA). Enhanced chemiluminescence (ECL) was purchased from GE Healthcare (Pittsburg, PA). Donkey anti-rabbit and donkey anti-mouse horseradish peroxidase (HRP) were purchased from Jackson ImmunoResearch (West Grove, PA). AMV reverse transcriptase was purchased from Promega (Madison, WI). SYBR Green PCR Master mix was purchased from Thermo Fisher Scientific, Inc. (Waltham, MA, USA). EDTA-free complete mini Protease Inhibitor Cocktail was purchased from Roche (Indianapolis, IN). NuPAGE Bis-Tris precast gels and Phosphate buffered saline (PBS) were purchased from Life Technologies (Grand Island, NY). Bicinchoninic acid (BCA) protein assay kit was obtained from Thermo Scientific (Rockford, IL). ProSignal Blotting Film was purchased from Genesee Scientific (El Cajon, CA). Ethyl alcohol (190 proof) was purchased from VWR (Radnor, PA).
9
Western Blot Analysis of Cellular Signaling
10
Protein Expression Analysis in Cells
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Whole-cell lysates were prepared using radioimmunoprecipitation assay (RIPA) buffer, immunoblotted as described,32 (link),33 (link) and analyzed using the following primary antibodies: anti-FAK, anti-phospho-p27 (S10), anti-p27, anti-c-Src, anti-β-actin, anti-α-tubulin, and anti-GAPDH (glyceraldehyde 3-phosphate dehydrogenase; Santa Cruz Biotechnology, Santa Cruz, CA, USA); anti-phospho-c-Src (Y416), anti-phospho-ERK1/2, anti-ERK1/2, anti-phospho-STAT3 (Y705), and anti-STAT3 (Cell Signaling, Danvers, MA, USA); anti-phospho-FAK (Y397) (Abcam, Cambridge, UK); anti-BMI1 (Millipore, Temecula, CA); anti-vimentin (Sigma, St. Louis, MO); anti-HA (Roche, Mannheim, Germany); and rabbit anti-human TM4SF57 (link) and rabbit anti-mouse TM4SF5, which was produced using a peptide-corresponding mouse TM4SF5 (amino acid residues 117–138; CLIDNKWDYHFQETEGAYLRND) by ProSci (Poway, CA, USA).
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