Phospho mkk3 mkk6

Manufactured by Cell Signaling Technology

Sourced in United States

Phospho-MKK3/MKK6 is a lab equipment product that detects the phosphorylation of the mitogen-activated protein kinase kinases MKK3 and MKK6. These kinases play a role in cellular signaling pathways.

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2 protocols using phospho mkk3 mkk6

Immunohistochemical Analysis of Bone Sections

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Dissected bones were fixed in 10% formalin and decalcified in formical-4^™ (Decal Chemical Corporation) for 3 days before embedding in paraffin wax [17 (link)]. Bone sections (3–4μm) were stained with Alcian blue 8GX and van Gieson [17 (link)]. For immunohistochemical analysis, dewaxed sections were treated with proteinase K for antigen retrieval, before staining with rabbit polyclonal phospho-p38 MAPK (cat # 4631) and phospho-MKK3/MKK6 (cat # 9231) antibodies (Cell Signalling Technology, Danvers, USA) using the ABC staining system (Santa Cruz Biotechnology, Santa Cruz, USA). Digital images were obtained using the Nanozoomer 2.0 Digital Pathology system (Hamamatsu Photonics, Welwyn Garden City, UK).

Esapa C.T., Piret S.E., Nesbit M.A., Loh N.Y., Thomas G., Croucher P.I., Brown M.A., Brown S.D., Cox R.D, & Thakker R.V. (2016). Mice with an N-Ethyl-N-Nitrosourea (ENU) Induced Tyr209Asn Mutation in Natriuretic Peptide Receptor 3 (NPR3) Provide a Model for Kyphosis Associated with Activation of the MAPK Signaling Pathway. PLoS ONE, 11(12), e0167916.

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Western Blot Analysis of Stress Signaling

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After transfections and/or treatment with chemicals or not, cells were lysed for the Western Blot assay. Bands were incubated with primary antibodies against SIRT1 (#2493),p16^INK4a (#4824), p21^WAF (#2947), p53 (#9282), MKK6 (#8550),MKK3 (#5674), Phospho-MKK3/MKK6 (#9236), Phospho-p38 MAP Kinase (#9211), p38 MAP Kinase (#9212), Phospho-Hsp27 (#2401), Phospho-MAPKAPK2 (#3007), Phospho-ATF2 (#5112) and actin (#8457) purchased from Cell Signaling Technology (Danvers, MA) and used at the recommended dilution for immunoblotting (1:1000). Raw data presenting full-length blots bands with molecular size markers are shown in Figure S9.

Simoncini S., Chateau A.L., Robert S., Todorova D., Yzydorzick C., Lacroix R., Ligi I., Louis L., Bachelier R., Simeoni U., Magdinier F., Dignat-George F, & Sabatier F. (2017). Biogenesis of Pro-senescent Microparticles by Endothelial Colony Forming Cells from Premature Neonates is driven by SIRT1-Dependent Epigenetic Regulation of MKK6. Scientific Reports, 7, 8277.

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