Guinea pig anti-insulin (Dako A0564, 1:500), and rabbit anti-glucagon (Millipore AB932, 1:500) antibodies for immunofluorescence were used on paraffin-embedded pancreata fixed in 4% paraformaldehyde (PFA). Plasma glucose levels were obtained with the One Touch glucometer (Bayer). Plasma and total pancreatic insulin measurements were measured by radioimmunoassay or ELISA as described (Millipore, Crystal Chem)40 (link). Plasma leptin was measured by ELISA (Peprotech) and adiponectin by radioimmunoassay (Millipore).
Ab932
Manufactured by Merck Group
The AB932 is a piece of laboratory equipment designed for DNA extraction and purification. It utilizes automated magnetic bead-based technology to efficiently isolate high-quality nucleic acids from a variety of sample types. The core function of the AB932 is to streamline the DNA extraction process, providing consistent and reliable results.
Automatically generated - may contain errors
Lab products found in correlation
Adiponectin, by Merck Group (2 mentions)
A0564, by Agilent Technologies (2 mentions)
Sc 9168, by Santa Cruz Biotechnology (1 mentions)
Axio imager m2 fluorescence microscope, by Zeiss (1 mentions)
Lsm 710 nlo, by Zeiss (1 mentions)
Alexa fluor 488 donkey anti mouse igg h l antibody, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 594 affinipure donkey anti guinea pig igg h l, by Jackson ImmunoResearch (1 mentions)
4 protocols using ab932
1
Quantification of Insulin and Glucagon
2
Immunohistochemical Staining of Pancreatic Islets
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After overnight fixation in 4% paraformaldehyde at 4 °C, the pancreases dissected from neonatal mice were putted in 10% sucrose for 4 h, 20% sucrose for 8 h, and 30% sucrose for 12 h, sequentially. The pancreases slices were first incubated at 4 °C with blocking buffer for 90 min, and then incubated overnight at 4 °C with anti-insulin (cat. #sc-9168, Santa Cruz, 1:250), anti-SST (cat. ab30788, Abcam, 1:500), or anti-glucagon (cat. #AB932, Millipore, 1:250) antibodies. After washing three times with PBS, the slices were incubated at 4 °C for 2.5 h with goat anti-rabbit antibody (cat. #A27034, Invitrogen) or goat anti-rat antibody (cat. #ab150158, Abcam). The slices were stained with 4′, 6′-diamidino-2-phenylindole and subjected to fluorescence microscopic analysis as previously described47 (link),48 (link).
3
Immunohistochemical Analysis of Pancreatic Islet Cells
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Tissues were fixed in 4% paraformaldehyde overnight at 4 °C. For frozen sections, the tissues were cryoprotected in 30% sucrose, embedded in Optimal Cutting Temperature (OCT) media (Thermo Fisher, 6502), and frozen on dry ice. For paraffin sections, the tissues were dehydrated with 30–100% ethanol and embedded in paraffin. The sections (6 μm thick) were probed with primary antibodies against insulin (1:500, Abcam, ab7842), NKX6.1 (1:100, DSHB, F55A12-c), GLUT2 (1:50, Santa Cruz Biotechnology, sc-518022), and glucagon (1:200, Millipore, AB932) followed by incubation with Alexa Fluor 594 AffiniPure Donkey Anti-Guinea Pig IgG (H + L) (Jackson ImmunoResearch, 706-585-148) and Alexa Fluor 488 Donkey Anti-Mouse IgG (H + L) antibody (Thermo Fisher, A21202) as the secondary antibodies. Images were acquired using a Zeiss Axioimager M2 fluorescence microscope or Zeiss LSM 710 NLO and DuoScan System.
4
Quantification of Insulin and Glucagon
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