primary antibodies against tra-1-60  (Novus Biologicals)

Journal: International Journal of Molecular Sciences

Article Title: Transcriptome Analysis of Mesenchymal Progenitor Cells Revealed Molecular Insights into Metabolic Dysfunction and Inflammation in Polycystic Ovary Syndrome

doi: 10.3390/ijms25147948

Figure Lengend Snippet: Characterization of induced pluripotent stem cells (iPSCs) and iPSC-derived mesenchymal progenitor cells (iPSCMs). ( a ) Alkaline phosphatase staining showing alkaline phosphatase activity in iPSCs (blue). ( b ) Immunostaining for the pluripotency markers NANOG (red) and TRA-1-60 (green) in iPSCs. NANOG was expressed in the nuclei, while TRA-1-60 was expressed on the cell surface. Nuclei were stained with Hoechst 33,342 (blue). ( c ) Hematoxylin and eosin (H&E) staining of teratomas derived from iPSCs. The markers show the sites of lineage-specific morphological characteristics, including neuroepithelial rosette (NR) for ectoderm, chondrocyte (C) for mesoderm, and intestinal or gut-like epithelium (I/E) for endoderm. ( d ) Bright-field image showing the fibroblast-like spindle morphology of iPSCMs. ( e ) Oil red O staining of lipid droplets in adipocytes derived from iPSCMs. ( f ) Alcian blue staining for acidic polysaccharides in chondrocytes derived from iPSCMs. ( g ) Alizarin Red S staining of calcium nodules in osteoblasts derived from iPSCMs. Scale bars, 100 mm.

Article Snippet: This experiment involved the use of primary antibodies against NANOG (Cell Signaling, Danvers, MA, USA) and TRA-1-60 (Millipore, Burlington, MA, USA).

Techniques: Derivative Assay, Staining, Activity Assay, Immunostaining

Journal: Stem Cell Reports

Article Title: Modeling G2019S-LRRK2 Sporadic Parkinson's Disease in 3D Midbrain Organoids

doi: 10.1016/j.stemcr.2019.01.020

Figure Lengend Snippet: Generation of Midbrain 3D Organoids from hiPSCs (A) Bright-field microscopy images of the stages of 3D organoid generation for 2 months. Scale bars, 200 μm. (B) qRT-PCR analysis of a pluripotency marker ( OCT4 ), a neural progenitor marker ( SOX1 ), and dopaminergic neuronal markers ( TH and VMAT2 ) at different time points. Data represent the mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.01 by ANOVA. (C) Immunofluorescence for MAP2, MASH1, TUJ1, VMAT2, TH, DAT, and GIRK2 to confirm the presence of midbrain dopaminergic neurons on day 60. Scale bars, 50 μm. (D) Schematic image of midbrain development. The marginal zone (MZ) contains midbrain dopaminergic neurons that differentiate from radial glia cells in the ventricular zone (VZ). (E) Percentage of MAP2/MASH1-positive cells in the MZ- and VZ-like zones at day 60. (F) Gene expression profiling using qRT-PCR from 1 to 45 days. Red and green represent higher and lower gene expression levels, respectively; n = 3 per sample. (G) Fluorescence-activated cell sorting analysis of synapsin-RFP-positive cells from midbrain 3D organoids. (H and I) KCL-induced dopamine levels in midbrain 3D organoids using liquid chromatography-mass spectrometry analysis. Data represent the mean ± SEM. ∗∗ p < 0.01 by ANOVA. (J) Gene set enrichment analysis of the microarray data from midbrain 3D organoids compared with that of 2D cultures.

Article Snippet: The hiPSCs were immunostained using primary antibodies against OCT4 (Santa Cruz Biotechnology, sc-5279), SOX2 (R&D Systems, MAB2018), NANOG (Bethyl Laboratories, A300-397a), SSEA1 (Santa Cruz, sc-21702), and TRA 1–60 (Millipore, MAB4360), and appropriate fluorescent secondary antibodies (Invitrogen).

Techniques: Microscopy, Quantitative RT-PCR, Marker, Immunofluorescence, Gene Expression, Fluorescence, FACS, Liquid Chromatography, Mass Spectrometry, Microarray