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Guinea pig anti insulin

Manufactured by Biosynth
Sourced in United States

Guinea pig anti-insulin is a laboratory reagent used for the detection and quantification of insulin in biological samples. It is a polyclonal antibody raised in guinea pigs against insulin, a hormone produced by the pancreas that regulates blood sugar levels. This product can be used in various immunoassay techniques, such as ELISA, to measure insulin concentrations in research and clinical settings.

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2 protocols using guinea pig anti insulin

1

Immunofluorescent Imaging of Human Islet Cells

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Human islets were trypsinized so that a cell suspension was obtained, and then transferred to a clearly defined area of poly-lysine glass slides using a Cytospin 4 centrifuge (Thermo Scientific, Waltham, MA, USA). Thereafter cells were fixed with 4% (w/v) paraformaldehyde for 5–10 min at room temperature, blocked for 30 min using 2% BSA, permeabilized with 0.1% Tris-NaCl for 10 min, and stained overnight with the following primary antibodies: rabbit anti-GDF15 (Abcam, 1:250), guinea pig anti-insulin (Fitzgerald, Sudbury, MA, USA, 1:250), and mouse anti-glucagon (Thermo Fischer Scientific, 1:1000). All slides were washed with phosphate-buffered saline (PBS) and, prior to counterstaining, with diluted secondary antibodies: ALEXA Fluor 594 donkey anti-rabbit (Jackson, bar Harbor, ME, USA, 1:250), ALEXA Fluor 488 donkey anti-guinea pig (Jackson, 1:250), and ALEXA Fluor 594 goat anti-mouse (Life technologies, Carlsbad, CA, USA, 1:250) for 1 h at room temperature. Cells were washed with PBS and then mounted with Biotium’s (Freemont, CA, USA) EverBrite™ mounting medium containing 4’,6-diamidino-2-phenylindole (DAPI) for nuclei staining. Images were taken and analyzed with a Nikon Eclipse C1/TE2000-U microscopy (Nikon, Konan, Tokyo, Japan).
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2

Immunostaining for Insulin Expression

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Adjacent sections of frozen in vitro autoradiography were immunostained for insulin expression. Sections were fixed in acetone (4 °C) for 5 min, allowed to dry for 15 min, and then washed in PBS and Dako washing buffer. Donkey serum (3% in PBS) was added for blocking and incubated for 20 min at RT, followed by overnight incubation with guinea pig anti-insulin 1:400 (Fitzgerald, Acton, MA, USA). Cat No. 20-IP30) at 4 °C. The next day, the sections were first washed in PBS 2 × 5 min and 1 × Dako washing buffer. Alexa 488-conjugated donkey anti-guinea pig secondary antibody (1:300, Jackson ImmunoResearch Laboratories, West Grove, PA, USA). Code No. 706–545-148) were added to the sections for 50 min at RT. Afterwards, the sections were rinsed 3 × 5 min PBS and then incubated with DAPI for 10 min to achieve a background. Finally, the sections were washed 2 × PBS and mounted in fluorescence mounting medium on protective glass.
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