Pancreases from each group were excised and processed as described previously [27] (link). Sections were cut at a thickness of 5 µm, dewaxed in xylene, and rehydrated in a graded ethanol series. The sections were incubated in PBS containing 3% BSA for 1 h at room temperature to block non-specific binding and then with the following antibodies in PBS containing 3% BSA overnight at 4°C: anti-glucagon receptor (1∶100, bs-3945R; Bioss, Woburn, MA), anti-insulin (1∶100, I2018; Sigma, Madrid, Spain), anti-glucagon (1∶100, G2654; Sigma, Madrid, Spain), and anti 11b-HSD1 (1∶50, sc-20175; Santa Cruz Biotechnology). After washing, the sections were incubated with the corresponding secondary antibodies (1∶1000, Alexa Fluor; Molecular Probes, Leiden, The Netherlands) for 1 h at room temperature. Hoechst 33342 (Life Technologies, Madrid, Spain) was used for nuclear staining. Images were captured using a Leica TCS SP2 AOBS spectral confocal microscope.
Sc 20175
Manufactured by Santa Cruz Biotechnology
Sourced in Spain
Sc-20175 is a lab equipment product offered by Santa Cruz Biotechnology. It is designed for use in various scientific applications. The core function of this product is to facilitate specific laboratory procedures and experiments.
Automatically generated - may contain errors
Lab products found in correlation
Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Anti insulin, by Merck Group (1 mentions)
G2654, by Merck Group (1 mentions)
Anti glucagon, by Merck Group (1 mentions)
Tcs sp2 aobs spectral confocal microscope, by Leica (1 mentions)
I2018, by Merck Group (1 mentions)
Hoechst 33342, by Thermo Fisher Scientific (1 mentions)
Hybond ecl, by Cytiva (1 mentions)
Sc 12763, by Santa Cruz Biotechnology (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
2 protocols using sc 20175
1
Immunohistochemical Analysis of Pancreatic Cell Types
2
Protein Analysis in Adipose Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Protein contents of: fatty acid synthase (FAS (ab150508, Abcam)); glucocorticoid receptor alpha (GR-α (sc-12763, Santa Cruz Biotechnology)); 11B-hydroxysteroid dehydrogenase type 1 (11B-HSD1 (sc-20175, Santa Cruz Biotechnology)) were evaluated by Western blotting in the VAT (epididymal fat sample); neuropeptide Y (NPY (N9525, Sigma-Aldrich)) was evaluated by Western blotting in the ARC. Briefly, tissues were homogenized on RIPA buffer (50 mM Tris-HCl (pH 7.4), 1% NP-40, 150 mM NaCl, 1 mM EDTA, 1 mM PMSF, 1 mM Na 3 VO 4 , 1 mM NaF) and protease inhibitor cocktail from F. Hoffmann-La Roche Ltd (Basel, Switzerland). Homogenates were centrifuged for 5 min (1120 g, 4°C). Protein concentration in supernatants was determined using the Pierce BCA Protein Assay Kit from Thermo Fisher Scientific. Then, homogenates were analyzed by SDS-PAGE using 10 mg total protein. Samples were electroblotted onto Hybond ECL nitrocellulose membranes from Amersham Pharmacia Biotech (London, UK). Membranes were incubated with Trisbuffered saline (TBS) containing 2% albumin for 90 min.
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