Human PDX-1/IPF1 Antibody

Detection of Human PDX‑1/IPF1 by Western Blot. Western blot shows lysates of beta TC-6 mouse beta cell insulinoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human PDX-1/IPF1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2419) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for PDX-1/IPF1 at approximately 45 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

Detection of Mouse PDX‑1/IPF1 by Simple Western^TM. Simple Western lane view shows lysates of beta TC-6 mouse beta cell insulinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for PDX-1/IPF1 at approximately 48 kDa (as indicated) using 10 µg/mL of Goat Anti-Human PDX-1/IPF1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2419) 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.

PDX‑1/IPF1 in BG01V Human Embryonic Stem Cells. PDX-1/IPF1 was detected in immersion fixed BG01V human embryonic stem cells differentiated into pancreatic progenitor cells using Goat Anti-Human PDX-1/IPF1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2419) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 493-conjugated Anti-Goat IgG Secondary Antibody (green; Catalog # NL003) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.

PDX‑1/IPF1 in Human Pancreatic Cancer Tissue. PDX-1/IPF1 was detected in immersion fixed paraffin-embedded sections of human pancreatic cancer tissue using Goat Anti-Human PDX-1/IPF1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2419) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei in cancer cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Human PDX-1/IPF1 by Immunocytochemistry/Immunofluorescence Differentiation of MODY3-iPSCs into pancreatic beta cells.(A) scheme of 6 step differentiation protocol (B), expression pattern of HNF1A mRNA during differentiation process, (C) Phase-contrast image of spheroids of differentiated MODY3-iPS-beta cells, (D, E) Immunocytochemistry of differentiated MODY3-iPS-beta cells for pancreatic beta cell markers (D) C-peptide, PDX1, NKX6.1 and DAPI staining, (E) HNF1A and DAPI staining. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31145732), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of PDX‑1/IPF1 in Human Pancreas. Formalin-fixed paraffin-embedded tissue sections of human pancreas were probed for PDX-1 mRNA (ACD RNAScope Probe, catalog #437088; Fast Red chromogen, ACD catalog # 322750). Adjacent tissue section was processed for immunohistochemistry using goat anti-human PDX-1 polyclonal antibody (R&D Systems catalog # AF2419) at 1ug/mL with overnight incubation at 4 degrees Celsius followed by incubation with anti-goat IgG VisUCyte HRP Polymer Antibody (Catalog # VC004) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to islet cells and exocrine glands.

Detection of Human Human PDX-1/IPF1 Antibody by Flow Cytometry GP2 and CD142 comparative profiles. a–j Flow cytometry analyses of H1-derived day 13 PP cultures obtained using 10 mM nicotinamide (a–e, optimal differentiation) and 3.3 mM nicotinamide (f–j, suboptimal differentiation) during stage 4 of differentiation. Cells were stained with anti-GP2 and anti-CD142 and then fixed and stained for the intracellular markers PDX1 and NKX6-1. IgG controls are shown on the left of each panel. Green boxes highlight the low percentage of GP2+/PDX− cells (c–h), red box highlights the presence of CD142+PDX1− cells (j). Abbreviations: GP2, pancreatic secretory granule membrane major glycoprotein 2; CD142, tissue factor; NKX6-1, NK6 homeobox 1; PDX1, pancreatic, and duodenal homeobox 1 Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28835709), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Human PDX-1/IPF1 Antibody by Flow Cytometry GP2 and CD142 comparative profiles. a–j Flow cytometry analyses of H1-derived day 13 PP cultures obtained using 10 mM nicotinamide (a–e, optimal differentiation) and 3.3 mM nicotinamide (f–j, suboptimal differentiation) during stage 4 of differentiation. Cells were stained with anti-GP2 and anti-CD142 and then fixed and stained for the intracellular markers PDX1 and NKX6-1. IgG controls are shown on the left of each panel. Green boxes highlight the low percentage of GP2+/PDX− cells (c–h), red box highlights the presence of CD142+PDX1− cells (j). Abbreviations: GP2, pancreatic secretory granule membrane major glycoprotein 2; CD142, tissue factor; NKX6-1, NK6 homeobox 1; PDX1, pancreatic, and duodenal homeobox 1 Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28835709), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Human PDX-1/IPF1 Antibody by Flow Cytometry MAC-sorted GP2+ cells give rise to ‘ beta -like’ cells in vitro. a Flow plots showing the GP2 profile at day 13 of differentiation of H9 cells. Cells were analyzed either before MACS sorting (presort), after GP2 enrichment from a positive selection column (GP2+) or in the flow through from a depletion column. Top right plot shows NKX6-1 and PDX1 expression by flow cytometry in day 13 unsorted (presort) H9 cells. b Following MACS sorting for GP2 at day 13, cells were cultured generate beta -like cells up to day 23. Representative flow cytometry plots of NKX6-1 and C-PEPTIDE (CPEP) expression at day 23 of differentiation from either unsorted (PRESORT), enriched for GP2 using a MACS positive selection column (GP2+) or in the flow through cell population (Flow-). The bar graph shows the average percentage of NKX6-1+/C-PEPTIDE+ cells at Day 23. N = 4, error bars indicate s.e.m. **p < 0.01, One-way ANOVA. c Model depicting the in vivo and in vitro equivalent of the human multipotent pancreatic progenitor (MPC) expressing PTF1A/GP2/NKX6-1. The MPC residing at the tip of the developing human pancreas has the potential to develop into acinar (PTF1A+/GP2+) and ductal/endocrine (NKX6-1+/GP2−) progenitors. Abbreviations: GP2, pancreatic secretory granule membrane major glycoprotein 2; D13, day 13; NKX6-1, NK6 homeobox 1; PDX1, pancreatic and duodenal homeobox 1; PTF1A, pancreas specific transcription factor 1a Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28835709), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Human PDX-1/IPF1 Antibody by Flow Cytometry GP2 and CD142 comparative profiles. a–j Flow cytometry analyses of H1-derived day 13 PP cultures obtained using 10 mM nicotinamide (a–e, optimal differentiation) and 3.3 mM nicotinamide (f–j, suboptimal differentiation) during stage 4 of differentiation. Cells were stained with anti-GP2 and anti-CD142 and then fixed and stained for the intracellular markers PDX1 and NKX6-1. IgG controls are shown on the left of each panel. Green boxes highlight the low percentage of GP2+/PDX− cells (c–h), red box highlights the presence of CD142+PDX1− cells (j). Abbreviations: GP2, pancreatic secretory granule membrane major glycoprotein 2; CD142, tissue factor; NKX6-1, NK6 homeobox 1; PDX1, pancreatic, and duodenal homeobox 1 Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28835709), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Human PDX-1/IPF1 Antibody by Flow Cytometry GP2 and CD142 comparative profiles. a–j Flow cytometry analyses of H1-derived day 13 PP cultures obtained using 10 mM nicotinamide (a–e, optimal differentiation) and 3.3 mM nicotinamide (f–j, suboptimal differentiation) during stage 4 of differentiation. Cells were stained with anti-GP2 and anti-CD142 and then fixed and stained for the intracellular markers PDX1 and NKX6-1. IgG controls are shown on the left of each panel. Green boxes highlight the low percentage of GP2+/PDX− cells (c–h), red box highlights the presence of CD142+PDX1− cells (j). Abbreviations: GP2, pancreatic secretory granule membrane major glycoprotein 2; CD142, tissue factor; NKX6-1, NK6 homeobox 1; PDX1, pancreatic, and duodenal homeobox 1 Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28835709), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Human PDX-1/IPF1 Antibody by Flow Cytometry GP2 and CD142 comparative profiles. a–j Flow cytometry analyses of H1-derived day 13 PP cultures obtained using 10 mM nicotinamide (a–e, optimal differentiation) and 3.3 mM nicotinamide (f–j, suboptimal differentiation) during stage 4 of differentiation. Cells were stained with anti-GP2 and anti-CD142 and then fixed and stained for the intracellular markers PDX1 and NKX6-1. IgG controls are shown on the left of each panel. Green boxes highlight the low percentage of GP2+/PDX− cells (c–h), red box highlights the presence of CD142+PDX1− cells (j). Abbreviations: GP2, pancreatic secretory granule membrane major glycoprotein 2; CD142, tissue factor; NKX6-1, NK6 homeobox 1; PDX1, pancreatic, and duodenal homeobox 1 Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28835709), licensed under a CC-BY license. Not internally tested by R&D Systems.