New Reagents, Publications and Data
We are intensifying our focus on Vision System Research with the addition of new antibodies. We are offering 50 USD off. They include: PSD-95/SAP90, Rhodopsin (A531), Rhodopsin (B630) and select Calcium (Ca2+) Signaling-Binding antibodies.
Image: Rhodopsin staining of pig retinal sections (green) and counter-stained with NF-M (red) and DNA (blue). Rhodopsin is most abundant in the outer segments of retina (OS), NF-M is abundant in the optic nerve fiber layer (ONFL), but seen in processes and neurons in other regions also. Other layers are pigmented epithelium (PE), outer and inner nuclear layers (ONL, INL), outer and inner plexiform layers (OPL, IPL) and ganglion cell layer (GCL). Inset: Bovine retinal extracts blotted with rhodopsin. Protocols on data-sheet.
Our ability to effectively serve researchers is confirmed by the growing parade of publications. I would like to highlight a recent publication by Dr. Sal Salvatore and his colleagues. It features use of our Shank 1a antibody. They are the first to show Shank 1 expression in the mammalian retina revealing Shank 1 immunoreactivity within both synaptic layers of the retina: Salvatore L. Stella Jr, Alejandro Vila, Albert Y. Hung, Michael E. Rome, Uyenchi Huynh, Morgan Sheng, Hans-Juergen Kreienkamp, Nicholas C. Brecha. Association of Shank 1A Scaffolding Protein with Cone Photoreceptor Terminals in the Mammalian Retina. PLoS ONE 7(9): e43463. doi:10.1371/journal.pone.0043463.
Images: Shank 1A immunoreactivity is in both the inner plexiform layer (IPL) and outer plexiform layer (OPL) of the mouse YFP-16 line retina. A–C: A. Image of a retinal section immunostained for Shank 1A. B. Mouse YFP-16 line vertical retinal section. C. Shank 1A (red) immunolabeling and YFP (yellow). Shank1A expression is restricted to the OPL and IPL. A regular pattern of Shank 1A immunolabeling appears in the OPL, which is indicative of cone photoreceptor terminals. D–E: High magnification zoom of the OPL demonstrates that Shank 1A puncta (red) are distal to the dendrite tips (yellow) of YFP labeled cone bipolar cells, suggesting that Shank 1A is expressed presynaptic to the YFP cone bipolar cell dendrite. G–L: High magnification zoom of the IPL demonstrates that Shank 1A puncta are likely expressed postsynaptically to bipolar cell terminals. G. Shank 1A immunoreactive puncta. H. YFP labeled neurons and processes within the IPL region. I. PKCα labeled rod bipolar cell axons and terminals. J. Combined Shank 1A (red) and PKCα (blue) immunolabeling illustrate that shank 1A puncta are postsynaptic to rod bipolar cell terminals in the IPL. K. Combined Shank 1A (red) immunolabeling and YFP (yellow) in the IPL demonstrate that Shank 1A puncta are postsynaptic to cone bipolar cell terminals in the IPL. L. Combined triple fluorescent image of Shank 1A (red), PKCα (blue), and YFP (yellow) in the IPL. OPL = outer plexiform layer, INL = inner nuclear layer, IPL = inner plexiform layer, and GCL = ganglion cell layer. Scale bars = 10 µm. doi:10.1371/journal.pone.0043463.g001
Diego C. Fernandez, Laura A. Pasquini, Damián Dorfman, Hernán J. Aldana Marcos, Ruth E. Rosenstein. Early Distal Axonopathy of the Visual Pathway in Experimental Diabetes. doi:10.1016/j.ajpath.2011.09.018
...a goat polyclonal anti–platelet-derived growth factor receptor-α (PDGFR-α) antibody (1:100; Neuromics, Edina, MN)
Raoul Torero Ibad, Jinguen Rheey, Sarah Mrejen, Valérie Forster, Serge Picaud, Alain Prochiantz, and Kenneth L. Moya. Otx2 Promotes the Survival of Damaged Adult Retinal Ganglion Cells and Protects against Excitotoxic Loss of Visual Acuity In Vivo. The Journal of Neuroscience, 6 April 2011, 31(14): 5495-5503; doi: 10.1523/JNEUROSCI.0187-11.2011...For antibody neutralization experiments, anti-Otx2 antibody (Neuromics) was dialyzed against PBS, and then Otx2 (25 ng in 500 mul) was incubated with anti-Otx2 (0.5 mug) in culture medium at 37C...
Hoon Shim, Chih-Ting Wang, Yen-Lin Chen, Viet Q. Chau, Kevin G. Fu, Jianqi Yang, A. Rory McQuiston, Rory A. Fisher, and Ching-Kang Chen. Defective Retinal Depolarizing Bipolar Cells (DBCs) in Regulators of G-protein Signaling (RGS) 7 and 11 Double Null Mice. JBC Papers in Press. Published on February 27, 2012 as Manuscript M112.345751. The latest version is at http://www.jbc.org/cgi/doi/10.1074/jbc.M112.345751...Animals were sacrificed by CO2 inhalation and the eyeballs were immediately enucleated. After removal of cornea and lens the resulting eyecups were immersionfixed in 4% paraformaldehyde in 1X PBS at room temperature for 15 minutes. This short fixation time ensured good mGluR6 and Gb5 signals at the OPL. After cryoprotection in 30% sucrose in 1X PBS, the eyecups were embedded in TBS (Richard Allan Scientific, Kalamazoo, MI), sectioned at 20μm thickness, and stained...
I will keep you posted on progress.
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