Mouse Gas1 Antibody

Gas1 in Mouse Embryo. Gas1 was detected in immersion fixed frozen sections of mouse embryo (13 d.p.c.) using Goat Anti-Mouse Gas1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2644) at 5 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 developing brain. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Detection of Mouse Gas1 by Immunocytochemistry/Immunofluorescence Immunofluorescent staining of E15.5 Wild type HoxAADD (a–d) and mutant Hoxaadd (Hoxa9,10,11−/−;Hoxd9,10,11−/−) (e–h) forelimbs. Arrowhead: radius, Arrow: ulna, the autopod is oriented to the left of the image. a and e: Six2 immunostaining, showing an increased expression in mutant chondrocytes. b and f: Gas1 staining, showing an increase in mutant limbs that is restricted to cells flanking chondrocytes, consistent with the inclusion of some perichondrial cells in the LCM samples. c and g: Lef1 staining, showing an absence of staining in mutant chondrocytes. d and h: Runx3 staining, showing an absence of staining in mutant chondrocytes Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/26186931), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse Gas1 by Immunocytochemistry/Immunofluorescence Immunofluorescent staining of E15.5 Wild type HoxAADD (a–d) and mutant Hoxaadd (Hoxa9,10,11−/−;Hoxd9,10,11−/−) (e–h) forelimbs. Arrowhead: radius, Arrow: ulna, the autopod is oriented to the left of the image. a and e: Six2 immunostaining, showing an increased expression in mutant chondrocytes. b and f: Gas1 staining, showing an increase in mutant limbs that is restricted to cells flanking chondrocytes, consistent with the inclusion of some perichondrial cells in the LCM samples. c and g: Lef1 staining, showing an absence of staining in mutant chondrocytes. d and h: Runx3 staining, showing an absence of staining in mutant chondrocytes Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/26186931), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse Gas1 by Western Blot Transfection of Hepa 1–6 cells with the growth-arrest specific 1 (Gas1) gene.(A) Anti-HA immunofluorescence of Hepa 1–6 cells transfected with pcDNA3/CAG-HAGas1 in the absence of detergents to preserve the integrity of membranes. Nuclei were counterstained with DAPI. (B) Double immunofluorescent staining anti-HA/anti-BrdU of Hepa 1–6 cells transfected with pcDNA3/CAG-HAGas1. (C) Cell cycle analysis of GFP-positive HEPA 1–6 cells after transfection with pIRES/GFP empty vector. 106 cells (areas indicated in the upper row) were sorted and subjected to cell cycle analysis after propidium iodide staining (lower row). (D) As (C), after transfection with pIRES/HAGas1. (E) Quantitation of the % of cells in the different stages of the cell cycle from the flow cytometry analysis. Experiments were done in triplicate. (F) Analysis of Ccne2 expression in Hepa 1–6 cells overexpressing Gas1. A representative RT-PCR showing Gas1 and Ccne2 expression in cells transfected with either empty pcDNA3/CAG (control) or pcDNA3/CAG-HAGas1 (HAGas1), 15 h after transfection. (G) qRT-PCR to determine CycE2 expression in cells as in (F). qRT-PCR was performed in triplicate from three independent experiments. Values were averaged and normalized to 18S rRNA. **, p<0.01. ***, p<0.001. In A and B the bar represents 11 μm. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/26161998), licensed under a CC-BY license. Not internally tested by R&D Systems.