Human/Mouse/Bovine Insulin Antibody

Insulin in betaTC‑6 Mouse Cell Line.

Insulin was detected in immersion fixed betaTC-6 mouse beta cell insulinoma cell line using Human/Mouse/Bovine Insulin Monoclonal Antibody (Catalog # MAB1417) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red; NL013) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Insulin in Human Pancreas.

Insulin was detected in immersion fixed paraffin-embedded sections of human pancreas using Rat Anti-Human/Mouse/Bovine Insulin Monoclonal Antibody (Catalog # MAB1417) at 0.5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rat IgG VisUCyte™ HRP Polymer Antibody (VC005). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and plasma membrane in islet cells. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of Insulin by Immunohistochemistry

Preservation of pancreatic islet insulin-producing beta cells from female NOD mice treated with AKS-107. Four to 6 week-old female NOD mice received i.p. injections of Vehicle or AKS-107 at 2 mg/kg twice weekly for 16 weeks (see Figure 8E for diabetes incidence) in which mice were sacrificed at 26 weeks of age for insulin- and C-peptide-specific cell staining via immunohistochemistry analyses of whole pancreas samples. Briefly, whole pancreata were harvested, formalin fixed, embedded, and sectioned such that longitudinal slices of pancreas were placed on slides. Sections were stained for insulin using a rat anti-insulin primary Ab (R & D systems, #MAB1417) and a donkey anti-rat IgG secondary Ab conjugated to Alexa488, and for leukocyte infiltration (insulitis) using Hematoxylin. Slides were scanned on an Aperio whole slide scanner (20x magnification), and then analyzed in a blinded fashion. One slide was analyzed per animal and for each animal the number of insulin islets per slide were counted. Mean insulin (A) and C-peptide (B) positive islets per slide per animal are shown (p, student’s t-test) beside representative images of positive (AKS-107-treated) and negative (Vehicle-treated) islet samples. Image collected and cropped by CiteAb from the following open publication (https://www.frontiersin.org/articles/10.3389/fimmu.2024.1367514/full), licensed under a CC-BY license. Not internally tested by R&D Systems.