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<br> The GLP-1 hormone also impacts areas of the brain that process how hungry a person is. So, GLP-1 as a natural hormone is lower potency than medication. Still, Johns doesn’t regret taking the medication. Images presented in Figure 4 (and Supplemental Figures S1 and S2), were taken on a Zeiss AxioImager Meta 510 confocal microscope (Carl Zeiss; Oberkochen, Germany). All sections were photographed using an Olympus BX51 reflected fluorescence system microscope (Olympus Denmark A/S; Ballerup, Denmark) equipped with darkfield, selective AMCA, FITC, and Texas red filters and a DP50 digital camera. Mice and rat pancreatic specimens were immersion fixed in 4% (w/v) paraformaldehyde in 0.1 M phosphate buffer, pH 7.4, for 24 hr at 4C, dehydrated in increasing concentrations of alcohol,  [http://www.riverbendadvisors.com/index.php?title=User:WoodrowWitt ColonBroom official] cleared in Estisol (Esti Chem A/S; Køge, Denmark), and embedded in paraffin in a Hypercenter 3000 tissue preparation machine (Leica Meyer Instruments; Houston, TX). Sections were deparaffinized in xylene and rehydrated in decreasing concentrations of alcohols. Sections were dehydrated in increasing concentrations of alcohol, cleared in 3M ammonium acetate, air dried, immersed in autoradiographic emulsion, and exposed for 4 weeks. When OXM is administered subcutaneously to overweight and obese subjects over a period of 4 weeks there is a significant reduction in body weight.<br><br><br><br> A subpopulation of the β cells were highly immunoreactive, whereas the great majority showed lower levels of expression (Figures 4A, 4B, and 4D), and there was no colocalization of the GLP-1R and  ColonBroom glucagon (Figures 4A, 4C-4F, and 4H) or somatostatin (Figures 4E, 4G, and 4H). Controls consisted of cross-absorption of the GLP-1R and insulin antibodies with either the KLH-conjugated GLP-1R peptide or human insulin. The isotype specific control antibody showed no unspecific staining reactivity. Fluor-labeled detection antibody is added to each well and incubated. Sections were incubated with guinea pig anti-human insulin, overnight at 4C, followed by FITC conjugated rabbit anti-guinea pig in TBS-T containing 7% (v/v) goat serum and/or 3% (v/v) species-specific serum/BSA for 30 min. Sections were cooled 10 min in the buffer, and endogenous peroxidase activity was blocked with 0.5% (v/v) H2O2 in 0.05 M TBS, pH 7.6, for 20 min. Sections were rinsed with TBS and incubated with Vectastain ABComplex for 30 min (Vector; Burlingame, CA), followed by preblocking in 0.1 M Tris buffer containing 0.15 M NaCl and 0.5% (w/v) Du Pont Blocking Reagent and biotinylated tyramide (TSA indirect; NEL 700) diluted 1:50 in amplification buffer (NEN; PerkinElmer Life Sciences, Boston, MA). After preincubation with TBS containing 10% (v/v) goat serum for 1 hr, the sections were incubated with rabbit anti-GLP-1R in TBS containing 7% (v/v) goat serum and 3% (v/v) species specific serum overnight at 4C. After washing, the sections were incubated with biotinylated goat anti-rabbit Ig in TBS containing 7% (v/v) goat serum and 3% (v/v) species specific serum (DakoCytomation) for  [https://fakenews.win/wiki/Discover_The_Benefits_Of_ColonBroom:_A_Comprehensive_Guide ColonBroom official] 1 hr.<br><br><br><br> To block endogenous biotin binding sites, sections were incubated with avidin blocking solution (DakoCytomation) for 10 min, washed in TBS, and incubated with the biotin blocking solution (DakoCytomation) for an additional 10 min.