fluorescent avidin biotin complex kit  (Vector Laboratories)

Journal: Cells

Article Title: Deletion of the Murine Ortholog of the Human 9p21.3 Locus Leads to Insulin Resistance and Obesity in Hypercholesterolemic Mice

doi: 10.3390/cells13110983

Figure Lengend Snippet: Chr4 Δ70/Δ70 does not affect pancreatic β-cell number but may reduce the cell proliferation rate already in young Ldlr −/− ApoB 100/100 mice. ( A ) Pancreatic islet size, ( B ) proportion of α- and β-cells, and ( C ) proliferation of mixed and ( D ) >10,000 μm 2 pancreatic islets in young Ldlr −/− ApoB 100/100 ( n = 5–7) and Chr4 Δ70/Δ70 mice ( n = 5–7). Primary antibody against glucagon was used as a marker for the pancreatic α-cells, and insulin ab was used for the β-cells. For visualization, fluorescent secondary antibodies were used. In the representative figures, glucagon positive cells appear in red and insulin in green. Nuclei were counterstained with DAPI (blue). Ki-67 antibody was used as a marker of cell proliferation and visualized with DAB. ( E ) For the islet function, insulin secretion of Ldlr −/− ApoB 100/100 ( n = 4) and Chr4 Δ70/Δ70 mice ( n = 4) was measured both in fasted state (0 min) and in response to 1 g/kg i.p. glucose at time points 15, 30, 60 and 90 min after the administration. Asterisk (*) indicates statistical significance. Difference in mean between Ldlr −/− ApoB 100/100 and Chr4 Δ70/Δ70 mice was measured by using t -test, and it was considered statistically significant when p < 0.05.

Article Snippet: Pancreatic α- and β-cells were immunostained with primary antibodies against glucagon (Dako A0565, Rabbit anti-human glucagon, Agilent, Santa Clara, CA, USA) and insulin (Dako A0564, Guinea pig anti-insulin, Agilent, Santa Clara, CA, USA) and appropriate fluorescent secondary antibodies (for glucagon, A21442, chicken anti-rabbit A594, Thermo Fisher Scientific, Waltham, MA, USA, and for insulin, BA-7000 Goat anti-guinea pig with A-2011 Fluorescein Avidin DCS, Vector laboratories, Newark, CA, USA).

Techniques: Marker