Whole mounts of mammary tissues from nonparous mice and from day 13 of pregnancy (p13) were fixed in Carnoy's mix, hydrated and stained with carmine alum. Paraffin sections were stained with H&E by standard methods. For immunostaining, primary antibodies were incubated overnight at 4°C (anti-H3K27me3, Millipore, 1:200; anti-EZH2, Cell Signaling, 1:200; anti-phosphorylated STAT5, Cell Signaling, 1:200; anti-WAP, Santa Cruz Biotechnology, 1:200; anti-E-cadherin, BD Biosciences, 1:100).
Anti phosphorylated stat5
Manufactured by Cell Signaling Technology
Anti-phosphorylated STAT5 is a laboratory reagent used for the detection and quantification of phosphorylated STAT5 protein in biological samples. STAT5 (Signal Transducer and Activator of Transcription 5) is a transcription factor that plays a critical role in cellular signaling pathways. Phosphorylation of STAT5 is a key regulatory event in various cellular processes.
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Lab products found in correlation
Anti phosphorylated stat3, by Cell Signaling Technology (2 mentions)
Anti stat5, by Cell Signaling Technology (2 mentions)
Anti wap, by Santa Cruz Biotechnology (1 mentions)
Anti ezh2, by Cell Signaling Technology (1 mentions)
Anti h3k27me3, by Merck Group (1 mentions)
Anti e cadherin, by BD (1 mentions)
Ecl kit, by Thermo Fisher Scientific (1 mentions)
Anti stat3, by Cell Signaling Technology (1 mentions)
Anti lc3, by Merck Group (1 mentions)
Anti hif 2α, by Novus Biologicals (1 mentions)
Anti β actin, by Cell Signaling Technology (1 mentions)
Laemmli sample buffer, by Bio-Rad (1 mentions)
Anti α tubulin, by Abcam (1 mentions)
Chlorpromazine, by Merck Group (1 mentions)
Anti flag antibody, by Merck Group (1 mentions)
Epidermal growth factor (egf), by Merck Group (1 mentions)
Filipin, by Merck Group (1 mentions)
Egf tritc, by Thermo Fisher Scientific (1 mentions)
Monoclonal anti actin, by Merck Group (1 mentions)
Alexa594 wga, by Thermo Fisher Scientific (1 mentions)
4 protocols using anti phosphorylated stat5
1
Mammary Tissue Whole Mount and Immunostaining
2
Immunohistochemical Analysis of STAT5 and STAT3 Phosphorylation in Mammary Tissues
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Mammary tissues from wild-type and mutant mice were harvested at L1 and fixed in 10% formalin and dehydrated in ethanol. Paraffin sections were stained with hematoxylin and eosin by standard methods (Histoserve). For immunofluorescence, unstained sections were incubated with the following antibodies overnight at 4°C: anti-phosphorylated STAT5 (Cell Signaling, 9314) and anti-phosphorylated STAT3 (Cell Signaling, 9145).
3
Western Blotting Protein Analysis Protocol
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Western blotting was performed as previously described (Liu et al., 2018 (link)). Briefly, cells were lysed in Laemmli sample buffer (cat# 161–0737, Bio-Rad) before SDS–PAGE. Primary antibodies used for western blotting were as follows: anti-FUNDC1 (home-made) (Liu et al., 2012a (link)), anti-LC3 (cat# L8918, Sigma), anti-TIMM23 (cat# 611222, Biosciences), anti-TOMM20 (cat# 612278, Biosciences), anti-STAT3 (cat# 9139, Cell Signaling Technology), anti-phosphorylated STAT3 (cat# 9134, Cell Signaling Technology), anti-STAT5 (cat# 94205, Cell Signaling Technology), anti-phosphorylated STAT5 (cat# 4322, Cell Signaling Technology), anti-HIF2α (cat# NB100-122, Novus Biologicals), anti-βACTIN (cat# 8H10D10, Cell Signaling Technology), and anti-αTUBULIN (cat# ab52866, Abcam). Horseradish peroxidase-conjugated secondary antibodies were used, and signals were detected using an ECL kit (cat# WP20005, Invitrogen).
4
Antibody Characterization and Endocytosis Assays
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Antibodies used were: monoclonal and polyclonal anti-FLAG antibodies from Sigma-Aldrich, polyclonal anti-Alix from Covalab, monoclonal anti-actin from Millipore, polyclonal antibodies against EGFR, Myc and EEA1 were from Santa Cruz, anti-GFP and anti-LAMP1 from Abcam, monoclonal anti-Rab11, monoclonal anti-clathrin and anti-GM130 from BD transduction, monoclonal anti-β1 (9EG7) was from BD Pharmingen, anti-STAT5 and anti- phosphorylated STAT5 were from Cell Signaling. Monoclonal anti-LBPA (6C4) and polyclonal anti-endophilin A antibodies were generous gifts from Jean Gruenberg (Geneva, Switzerland) and Pietro De Camilli (New Haven, United States) respectively. Monoclonal anti-IL2Rβ antibody (mAb 341) was described in39 (link). Secondary antibodies conjugated to Alexa Fluor 488, Alexa Fluor 594 and Alexa 594-WGA, as well as EGF-TRITC, CTxB-TRITC, Tf-TRITC and Dextran-TRITC were from Molecular Probes. Horseradish peroxidase-conjugated goat anti-mouse and anti-rabbit secondary antibodies were from Jackson Laboratories. Filipin, chlorpromazine, EGF, bafilomycin A1 were from Sigma-Aldrich. 6 nm BSA-Gold tracer was from Aurion (The Netherlands).
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