Immunofluorescent Pancreatic Cell Staining

Mouse pancreata were fixed in 4% paraformaldehyde and embedded with paraffin. Rehydrated 5 μm sections were stained with primary antibodies against insulin (dilution 1:1000; Dako, Santa Clara, CA, USA), somatostatin (dilution 1:300, GeneTex., Irvine, CA, USA), and glucagon (dilution 1:100; Abcam, Cambridge, UK) followed by secondary antibodies (dilution 1:500; anti-guinea pig Alexa Fluor 488, dilution 1:500; anti-mouse, Alexa Fluor 647, and dilution 1:500; anti-rabbit Alexa Fluor 546) as previously described (Vierra et al., 2015 (link)). Sections were imaged either with a Nikon Eclipse TE2000-U microscope or fluorescent ScanScope (Aperio).

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Nakhe A.Y., Dadi P.K., Kim J., Dickerson M.T., Behera S., Dobson J.R., Shrestha S., Cartailler J.P., Sampson L., Magnuson M.A, & Jacobson D.A. (2024). The MODY-associated KCNK16 L114P mutation increases islet glucagon secretion and limits insulin secretion resulting in transient neonatal diabetes and glucose dyshomeostasis in adults. eLife, 12, RP89967.

Publication 2024

somatostatin glucagon

Corresponding Organization :

Other organizations : Vanderbilt University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Expression of insulin, somatostatin, and glucagon proteins in mouse pancreatic tissue

control variables

Mouse pancreata were fixed in 4% paraformaldehyde and embedded with paraffin
Rehydrated 5 μm sections were stained with primary antibodies against insulin, somatostatin, and glucagon
Secondary antibodies (anti-guinea pig Alexa Fluor 488, anti-mouse Alexa Fluor 647, and anti-rabbit Alexa Fluor 546) were used
Sections were imaged either with a Nikon Eclipse TE2000-U microscope or fluorescent ScanScope (Aperio)

controls

No positive or negative controls were explicitly mentioned in the provided information

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