Immunoblotting analyses were performed as previously described [16 (link)]. Interleukin-17 (IL-17, Cell signaling Technology, Danvers, MA, USA), TGF-β1 (R&D Systems, Minneapolis, MN, USA), TGF-β inducible gene-h3 (βig-h3, Proteintech, Chicago, IL, USA), MnSOD (Abcam), Smad2/3 (Cell signaling Technology, Danvers, MA, USA), B-cell lymphoma-2 (Bcl-2, Santa Cruz Biotechnology), Bcl2-associated X (Bax, Delta Biolabs, Gilroy, CA, USA), and β-actin (Sigma) were detected with specific antibodies. Immunoblotting images were analyzed with an image analyzer (Quantity One, Bio-Rad Technical Service Department, Hercules, CA, USA). Optical densities were obtained using the VH group as 100% reference and normalized with β-actin.
βig h3
Manufactured by Proteintech
Sourced in United States
βig-h3 is a protein that regulates cell adhesion, differentiation, and migration. It is commonly used in research applications to study these cellular processes.
Automatically generated - may contain errors
3 protocols using βig h3
1
Immunoblotting Analysis of Signaling Proteins
2
Antibody Detection for Protein Analysis
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The antibodies used are as follows: PN, murine monoclonal (mAb) Stiny-1 (Adipogen, San Diego, CA) and rabbit polyclonal anti-PN ab14041 (Abcam, Cambridge, MA); βig-h3, murine mAb βig-h3 (Proteintech, Chicago, IL); γ-carboxyglutamyl (Gla) residues independent of protein context, murine mAb 3570 [18 (link)](Sekisui Diagnostics, Lexington, MA); fII, sheep polyclonal antibodies (Haematologic Technologies, Inc, Essex Junction, VT); and peroxidase conjugated anti-mouse, anti-Sheep, and anti-rabbit IgG (Jackson ImmunoResearch, West Grove, PA).
3
Immunohistochemical Analysis of Kidney Injury Markers
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Immunohistochemistry was performed as described previously. 18, 19 ED-1, osteopontin (OPN), transforming growth factor β1 (TGFβ1)-inducible gene-h3 (βig-h3), renin, 8-OHdG, and GLP-1R were detected in 4-μm tissue sections by incubating cells for 12 h with specific antibodies against kidney injury molecule-1 (KIM-1; Abcam, Cambridge, UK), neutrophil gelatinase-associated lipocalin (NGAL; Millipore, Billerica, MA, USA), ED-1 (AbD Serotec, Oxford, UK), OPN (obtained from the Developmental Studies hybridoma bank, University of Iowa, Iowa City, IA, USA), βig-h3 (Proteintech, Chicago, IL, USA), 8-OHdG (JaICA, Shizuoka, Japan), and GLP-1R (Abcam) at 4 °C. The KIM-1-, NGAL-, and βig-h3stained tissue sections were counted in 20 different fields in each section at × 200 magnification using color image analyzer (TDI Scope Eye Version 3.0 for Windows).
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