Tissue samples were sonicated in RIPA buffer (150 mM NaCl, 1% Triton X-100, 0.1% SDS, 0.5% Na deoxycholate, 50 mM Tris), and total protein was determined using BCA protein assays (Thermo-Scientific, Waltham, MA). Experiments were performed on the XCell II Blot Module (Invitrogen). From each sample, 20 μg of protein was loaded into a lane on a NuPage 4–12% Bis-Tris gel (Life Technologies, Carlsbad, CA), transferred to 0.2-μm polyvinylidene difluoride membrane (Bio-Rad, Hercules, CA), and processed with a blocking solution of 3% BSA and 5% nonfat dry milk in 1 × tris-buffered saline and Tween-20 (TBST). Rabbit anti-Cx36 (0.5 μl/ml; Invitrogen), rabbit anti-Cx43 (0.2 μl/ml; Invitrogen), and mouse anti-tubulin (1:10,000; Sigma-Aldrich, St. Louis, MO) were used as primary antibodies, and were visualized with horseradish peroxidase-conjugated anti-rabbit antibody (1:10,000; Invitrogen) or anti-mouse IgG antibody (1:10,000; Sigma-Aldrich). Signals were enhanced using enhanced chemiluminescence detection reagents (Thermo-Scientific) and detected on a BioSpectrum imaging system (UVP, Upland, CA). Densities were quantified using VisionWorks (UVP) and normalized relative to tubulin.
Rabbit anti cx36
Manufactured by Thermo Fisher Scientific
Rabbit anti-Cx36 is an antibody that specifically binds to the Connexin 36 (Cx36) protein. Cx36 is a gap junction protein found in various tissues, including the central nervous system. This antibody can be used for the detection and analysis of Cx36 expression in research applications.
Automatically generated - may contain errors
Lab products found in correlation
Enhanced chemiluminescence detection reagent, by Thermo Fisher Scientific (1 mentions)
Bca protein assay, by Thermo Fisher Scientific (1 mentions)
Xcell 2 blot module, by Thermo Fisher Scientific (1 mentions)
Anti mouse igg antibody, by Merck Group (1 mentions)
Horseradish peroxidase conjugated anti rabbit antibody, by Thermo Fisher Scientific (1 mentions)
Rabbit anti cx43, by Thermo Fisher Scientific (1 mentions)
Nupage 4 12 bis tris gel, by Thermo Fisher Scientific (1 mentions)
Visionworks, by Analytik Jena (1 mentions)
0.2 μm polyvinylidene difluoridemembrane, by Bio-Rad (1 mentions)
Mouse anti tubulin, by Merck Group (1 mentions)
Biospectrum imaging system, by Analytik Jena (1 mentions)
Sp8 scanning confocal microscope, by Leica (1 mentions)
Guinea pig anti insulin, by Agilent Technologies (1 mentions)
2 protocols using rabbit anti cx36
1
Comparative Analysis of Cx36 and Cx43 Protein Levels
2
Quantifying Islet Microenvironment Components
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Cx36 levels were quantified from images of islets co-stained with rabbit anti-Cx36 (Invitrogen) and guinea pig anti-insulin (Dako) antibody. Vasculature levels were quantified from images co-stained with rat anti-laminin β1 (Invitrogen) or rabbit anti-col IV and guinea pig anti-insulin (Dako). Innervation was quantified from images of tissue sections co-stained with rabbit anti-Tubb3 (BioLegend) and guinea pig anti-insulin (Dako). For vasculature and Cx36, eight Z-planes were taken 1 µm apart, and for innervation, five z-planes were taken 1 µm apart, on a Leica SP8 Scanning Confocal microscope using a 40× oil immersion objective (Cx36) or 20× objective (vasculature and innervation). Filtering and thresholding were applied to both channels for each islet, and the area of each staining was measured using Fiji’s analyze particles functions. The area of gap junctions, blood vessels, or innervation (marked by their respective antibody) was divided by the area of DAPI (for Cx36) or insulin (vasculature and innervation) for each islet.
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