Journal:

Article Title: Differentiation of spinal motor neurons from pluripotent human stem cells

doi: 10.1038/nprot.2009.127

Figure Lengend Snippet: Patterning of spinal cord progenitors. The hESC-derived neuroepithelial cells cultured without RA are positive for OTX2 (green) (a). When the cells are cultured in the presence of RA (100 nM) for 1 week, the majority of the cells express HOXB4 (red) (b). (c,d) Following a further week of culture, these progenitors retain the rostral (c) or caudal identity (d) when they differentiate to tubulin+ (Tuj1) (green) neurons. Both OTX2 and HOXB4 are represent as red in c and d. Bar µ 50 µm.

Article Snippet: list-behavior=unordered prefix-word= mark-type=disc max-label-size=0 Antibodies; refer to for details. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Antibody Isotype Source Cat. no. Dilution Oct4 Mouse IgG Santa Cruz Biotechnology sc-5279 1:1,000 Pax6 Mouse IgG DSHB NA 1:5,000 Sox1 Goat IgG R&D AF3366 1:1,000 Otx2 Goat IgG R&D AF1979 1:2,000 HoxB4 Rat IgG DSHB 112 anti-Hoxb4 1:50 Olig2 Goat IgG Santa Cruz Biotechnology SC-19969 1:500 MNR2 (HB9) Mouse IgG DSHB 81.5C10 1:50 ChAT Goat IgG Chemicon AB144P 1:200 βIII-tubulin Rabbit IgG Covance PRB-435P 1:5,000 Synapsin Rabbit IgG CALBIOCHEM 574777 1:250 α-bungarotoxin NA Molecular Probe B13423 1:500 Open in a separate window NA, not applicable.

Techniques: Derivative Assay, Cell Culture

Journal:

Article Title: Differentiation of spinal motor neurons from pluripotent human stem cells

doi: 10.1038/nprot.2009.127

Figure Lengend Snippet: Antibodies and probes used for determining the neural differentiation of hES cells.

Article Snippet: list-behavior=unordered prefix-word= mark-type=disc max-label-size=0 Antibodies; refer to for details. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Antibody Isotype Source Cat. no. Dilution Oct4 Mouse IgG Santa Cruz Biotechnology sc-5279 1:1,000 Pax6 Mouse IgG DSHB NA 1:5,000 Sox1 Goat IgG R&D AF3366 1:1,000 Otx2 Goat IgG R&D AF1979 1:2,000 HoxB4 Rat IgG DSHB 112 anti-Hoxb4 1:50 Olig2 Goat IgG Santa Cruz Biotechnology SC-19969 1:500 MNR2 (HB9) Mouse IgG DSHB 81.5C10 1:50 ChAT Goat IgG Chemicon AB144P 1:200 βIII-tubulin Rabbit IgG Covance PRB-435P 1:5,000 Synapsin Rabbit IgG CALBIOCHEM 574777 1:250 α-bungarotoxin NA Molecular Probe B13423 1:500 Open in a separate window NA, not applicable.

Techniques:

Journal: bioRxiv

Article Title: Xeno-free induced pluripotent stem cell-derived neural progenitor cells for in vivo applications

doi: 10.1101/2022.01.18.476253

Figure Lengend Snippet: A: Schematic representation of the analyzed cell types, time frame and the main marker proteins used for characterization. B: Principal component analysis of qPCR data for NPCs (3 clones, dark blue) iPSCs (3 clones, light blue), NPCs 21d after neural differentiation (clone 1, violet) and human total brain RNA. Each symbol illustrates data from RNA extracted from one cell culture well. C: Gene expression of pluripotency marker (NANOG and OCT4) and NPC marker (PAX6, SOX1, CXCR4, MSI1, NESTIN) in NPCs over the course of 15 passages, measured by qPCR. D: iPSCs (left panel) and NPCs at passage 7-10 (right panel) stained for Pax6, Nestin, Oct4 and DAPI. E: Flow cytometry analysis of iPSCs (upper row) and NPCs at different passages (lower rows) for Oct4, Pax6 and Sox1. Pie charts illustrate percentage positivity (light/dark blue) for the respective marker and cell type. n.t.: not tested Neural diff: NPCs after neural differentiation; CXCR4: CXC-motif chemokine receptor-4; MSI1: Musashi-1; Scale bars: 50μm. Significance of mean differences between the groups was assessed using Tukey’s HSD. Asterisks indicate significance: *** P < 0.001.

Article Snippet: To identify transplanted cells at different developmental stages, the following antibodies were used: Goat anti-human NANOG (R&D systems, 1:200), rabbit Oct-4A (Cell Signaling Technology, 1:200), mouse Anti-PAX6 monoclonal antibody (Thermo Fisher Scientific), mouse Anti-NeuN Antibody (Merck, #MAB377), rabbit Anti-Neurofilament 200 antibody (Merck, #N4142), rabbit Anti-BetaIII-Tubulin antibody (Abcam, #ab18207), mouse MAP2 monoclonal antibody (Thermo Fisher Scientific), rabbit Anti-S100b antibody (Thermo Fisher Scientific).

Techniques: Marker, Clone Assay, Cell Culture, Gene Expression, Staining, Flow Cytometry

Journal: PLoS ONE

Article Title: Apolipoprotein J/Clusterin in Human Erythrocytes Is Involved in the Molecular Process of Defected Material Disposal during Vesiculation

doi: 10.1371/journal.pone.0026033

Figure Lengend Snippet: (A, B) TEM immunogold localization of sCLU in RBCs membrane protrusions (A) and vesicles (ves) (B) collected from fresh units of stored RBCs (N = 2, young healthy donors). Solid or dashed arrows indicate sCLU immunogold localization at the periphery or the cytosol of the vesicles, respectively. (C) Representative immunoblot analysis of RBCs-derived purified vesicles (N = 2) probed with either polyclonal anti-sCLU or with monoclonal anti-Band 3 antibodies. Molecular weight markers are indicated to the right of the blot. Bars in (A), (B), 100 nm.

Article Snippet: The polyclonal antibody against CD59 was obtained from R&D Systems (AF 1987).

Techniques: Membrane, Western Blot, Derivative Assay, Purification, Molecular Weight

Journal: PLoS ONE

Article Title: Apolipoprotein J/Clusterin in Human Erythrocytes Is Involved in the Molecular Process of Defected Material Disposal during Vesiculation

doi: 10.1371/journal.pone.0026033

Figure Lengend Snippet: Purified RBCs membranes from healthy subjects (N = 6) were lysed in NP-40 and lysates were immunoprecipitated (IP) with polyclonal antibodies against sCLU, Band 3, stomatin or normal serum (control). Immunoprecipitates were immunoblotted (IB) under reducing conditions for sCLU (A 1 , upper panel), Band 3 (A 1 , middle panel), CD59 (A 1 , lower panel) and Hb (A 3 ); shown IPs are representatives from two independent experiments. (A 2 ) CLSM co-immunolocalization of the sCLU and Band 3 proteins at the RBCs plasma membrane. Cells were co-stained with anti-Band 3 monoclonal (green; upper panel) and anti-sCLU polyclonal antibodies (red; lower panel). Captured images were merged to reveal co-distribution sites (yellow; lower panel, arrows). Bars, 3 µm. (B) Anti-dinitrophenylhydrazone (DNP) immunoblotting of sCLU, Band 3, and control (IgGs) immunoprecipitates for the detection of co-immunoprecipitated carbonylated proteins (arrows) in 2,4-dinitrophenylhydrazine-modified (OX) or unmodified protein material.

Article Snippet: The polyclonal antibody against CD59 was obtained from R&D Systems (AF 1987).

Techniques: Purification, Immunoprecipitation, Control, Clinical Proteomics, Membrane, Staining, Western Blot, Modification

Journal: PLoS ONE

Article Title: Apolipoprotein J/Clusterin in Human Erythrocytes Is Involved in the Molecular Process of Defected Material Disposal during Vesiculation

doi: 10.1371/journal.pone.0026033

Figure Lengend Snippet: Erythrocytic sCLU localizes at both sides of the plasma membrane in association with non-cytoskeletal areas, as well as in the cytosol (see also, Antonelou et al., accompanying paper). At the intracellular side of the RBCs membrane sCLU may bind Band 3, Hb and/or other cytoskeleton-free membrane portions. On the other hand, the sCLU that localizes at the extracellular side of the RBCs membrane can attach to membrane by binding to Band 3, CD59, plasma membrane IgGs or to an currently unknown sCLU-specific receptor. Physiological in vivo or ex vivo RBCs senescence (1) is associated with cytosol, cytoskeleton and membrane structural alterations, including Band 3 modifications, increased membrane binding of IgGs, proteolysis, protein aggregation and increased oxidation defects. Vesiculation (2), a self-protective mechanism of mammalian erythrocytes, removes oxidized proteins and aggregates from both plasma membrane and cytosol thereby postponing the untimely elimination of otherwise healthy erythrocytes. This process takes place through the entire in vivo or ex vivo lifespan of RBCs and is functionally connected to the release of sCLU-, Band 3-, CD59-, Hb- and IgGs-containing vesicles. We propose that vesicular sCLU by following its membrane linkers (e.g. Band 3) or other unknown cytosolic interacting proteins assists via its chaperone function in the disposal of non-functional or death signalling effective material from RBCs.

Article Snippet: The polyclonal antibody against CD59 was obtained from R&D Systems (AF 1987).

Techniques: Clinical Proteomics, Membrane, Binding Assay, In Vivo, Ex Vivo, Functional Assay

Journal: Stem cell research

Article Title: Generation of two human iPSC lines with Exon 3 mutations in BCL2-Associated Athanogene 3 ( BAG3 ) from dilated cardiomyopathy patients

doi: 10.1016/j.scr.2023.103019

Figure Lengend Snippet: Reagents details.

Article Snippet: Differentiation Markers (Ectoderm) , Goat anti-OTX2 , 1:200 , R and D Systems Cat# AF1979, RRID: AB_2157172.

Techniques: Immunocytochemistry, Staining