Mouse Pref-1/DLK1/FA1 Antibody

Detection of Mouse Pref‑1/DLK1/FA1 by Western Blot. Western blot shows lysates of 3T3-L1 mouse embryonic fibroblast adipose-like cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Mouse Pref-1/DLK1/FA1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8277) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Pref-1/DLK1/FA1 at approximately 45-60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Pref‑1/DLK1/FA1 in 3T3‑L1 Mouse Cell Line by Flow Cytometry. 3T3-L1 mouse embryonic fibroblast adipose-like cell line was stained with Goat Anti-Mouse Pref-1/DLK1/FA1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8277, filled histogram) or control antibody (Catalog # AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107).

Pref‑1/DLK1/FA1 in Mouse Embryo. Pref-1/DLK1/FA1 was detected in immersion fixed frozen sections of mouse embryonic lung using Goat Anti-Mouse Pref-1/DLK1/FA1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8277) at 1 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 cytoplasm. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Detection of Mouse Pref-1/DLK1/FA1 by Immunohistochemistry Dlk1 imprinting & expression in the developing pituitary gland from the endogenous locus & TGDlk1-70C transgene.(A) Cross used to generate embryos & postnatal animals in the study. Males inheriting the deleted allele from the mother (maternal Dlk1tm1Srbpa/+ heterozygotes or MATs) were crossed to females hemizygous for the TGDlk1-70C transgene (WT-TG), generating 4 genotypes, WT, WT-TG, paternal Dlk1+/tm1Srbpa heterozygotes (PATs) & mice inheriting a deleted paternal allele & the transgene (PAT-TG). (B) Schematic showing the known splice variants of Dlk1, A-D. Splicing occurs internally in exon 5 of the Dlk1 gene. Dlk1-A & B retain an extracellular cleavage domain (TACE), in Dlk1-C & D this region is spliced out. All versions contain a single pass transmembrane domain (TM). Red arrows indicate location of primers used in (C). (C) Semi-quantitative PCR on embryonic day (E) 18.5 whole pituitary glands from the 4 genotypes shown in (A). Top – primers amplify the exon 4–5 region of Dlk1 & can distinguish splice variants based on size. Bottom – alpha-tubulin (Tuba1a) was amplified as a loading control on each sample. (D) In-situ hybridisation for Dlk1 in the developing pituitary gland from E9.5 to E18.5 in the 4 genotypes shown in (A). Dlk1 expression is indicated by purple staining. Scale bars show 100 µm (E9.5 & E13.5, sagittal sections) & 200 µm (E15.5 & E18.5, frontal sections). (E) Immunohistochemistry (IHC) for DLK1 on frontal sections at E18.5 & postnatal day 21 (P21), counterstained with DAPI. Scale bars = 50 µm.Figure 2—source data 1.Zipped file containing the source data for Figure 2D -gels with cropped areas highlighted & original gel images. Zipped file containing the source data for Figure 2D -gels with cropped areas highlighted & original gel images. Image collected & cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37589451), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse Pref-1/DLK1/FA1 by Immunohistochemistry Dlk1 imprinting & expression in the developing pituitary gland from the endogenous locus & TGDlk1-70C transgene.(A) Cross used to generate embryos & postnatal animals in the study. Males inheriting the deleted allele from the mother (maternal Dlk1tm1Srbpa/+ heterozygotes or MATs) were crossed to females hemizygous for the TGDlk1-70C transgene (WT-TG), generating 4 genotypes, WT, WT-TG, paternal Dlk1+/tm1Srbpa heterozygotes (PATs) & mice inheriting a deleted paternal allele & the transgene (PAT-TG). (B) Schematic showing the known splice variants of Dlk1, A-D. Splicing occurs internally in exon 5 of the Dlk1 gene. Dlk1-A & B retain an extracellular cleavage domain (TACE), in Dlk1-C & D this region is spliced out. All versions contain a single pass transmembrane domain (TM). Red arrows indicate location of primers used in (C). (C) Semi-quantitative PCR on embryonic day (E) 18.5 whole pituitary glands from the 4 genotypes shown in (A). Top – primers amplify the exon 4–5 region of Dlk1 & can distinguish splice variants based on size. Bottom – alpha-tubulin (Tuba1a) was amplified as a loading control on each sample. (D) In-situ hybridisation for Dlk1 in the developing pituitary gland from E9.5 to E18.5 in the 4 genotypes shown in (A). Dlk1 expression is indicated by purple staining. Scale bars show 100 µm (E9.5 & E13.5, sagittal sections) & 200 µm (E15.5 & E18.5, frontal sections). (E) Immunohistochemistry (IHC) for DLK1 on frontal sections at E18.5 & postnatal day 21 (P21), counterstained with DAPI. Scale bars = 50 µm.Figure 2—source data 1.Zipped file containing the source data for Figure 2D -gels with cropped areas highlighted & original gel images. Zipped file containing the source data for Figure 2D -gels with cropped areas highlighted & original gel images. Image collected & cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37589451), licensed under a CC-BY license. Not internally tested by R&D Systems.