Human Islet-1 Antibody

Detection of Human Islet‑1 by Western Blot. Western blot shows lysates of JOY6 human induced pluripotent stem cells undifferentiated and differentiatied into motor neurons. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Islet-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1837) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Islet-1 at approximately 42 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Islet‑1 in iPS2 Human Stem Cells. Islet-1 was detected in immersion fixed iPS2 human induced pluripotent stem cells differentiated to endocrine progenitor cells using Goat Anti-Human Islet-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1837) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI(blue). Specific staining was localized to the nucleus. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Islet‑1 in Human Pancreas. Islet‑1 was detected in immersion fixed paraffin-embedded sections of human pancreas using Goat Anti-Human Islet‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1837) at 1 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (VC004). 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 nuclei in islet cells. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of Human Islet‑1 by Simple Western^TM. Simple Western lane view shows lysates of JOY6 human induced pluripotent stem cells undifferentiated and differentiated into motor neurons, loaded at 0.2 mg/mL. A specific band was detected for Islet-1 at approximately 53 kDa (as indicated) using 10 µg/mL of Goat Anti-Human Islet-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1837) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of Mouse Islet-1 by Immunohistochemistry Efficiency of Isl1 ablation in stomachs of Isl1MCM/Del mutant mouse stomachs at E18.5. (A) Tamoxifen-inducible Cre recombinase excised DNA sequences flanked by two loxP sites. (B)Isl1 RNA levels were ablated in Isl1MCM/Del mutant stomachs as seen by semi-quantitative PCR. Isl1F/+mice showed a 592 base pair product whereas Isl1MCM/Del mice generated a 303 base pair product. (C) Isl1 was significantly down-regulated at the protein levels in Isl1MCM/Del mutant stomachs as shown by western blot. Expression of embryos at E11.5 was used as positive control. (D) Isl1 protein expression in Isl1F/+and Isl1MCM/Del embryonic pylorus. Isl1 expression was significantly reduced in Isl1MCM/Del embryonic stomachs, as seen by immunofluorescence. Images in Isl1F/+and Isl1MCM/Del were processed on the same slide and photographed at the same exposure. Enlarged images of the boxed areas are shown on the right side of the merged pictures. Yellow arrowheads show representative Isl1-positive cells, and white arrowheads show representative Isl1-negative cells. Yellow dotted lines mark the epithelial basement membrane. Scale bars: 50 μm. Image collected and cropped by CiteAb from the following publication (https://bmcbiol.biomedcentral.com/articles/10.1186/1741-7007-12-25), licensed under a CC-BY license. Not internally tested by R&D Systems.