Immunohistochemical Profiling of Retinal Cells

Retinal sections were stained with the following antibodies with our published protocols (34 ,35 (link)) Cone-arrestin (rabbit polyclonal, 1:1,000; Millipore), PKCα (rabbit polyclonal, 1:5,000; Sigma), recoverin (rabbit polyclonal, 1:2,000; Millipore), calbindin D28k (rabbit polyclonal, 1:1,000; Swant), neurofilament-RT97 (mouse monoclonal, 1:1,000; Millipore), parvalbumin (rabbit polyclonal, 1:3,000; Swant), synaptophysin (mouse monoclonal, 1:2,000; Millipore), RPE65 (mouse monoclonal, 1:1,000; Millipore), type IV collagen (rabbit polyclonal, 1:1,000; Millipore), MAB1281 (mouse monoclonal, 1:300; Millipore), nestin (rabbit polyclonal, 1:2,000; Millipore), GFAP (rabbit polyclonal, 1:1,000; Sigma), S100β (mouse monoclonal, 1:250; Sigma), Ki67 (rabbit polyclonal, 1:500; Millipore), TuJ1 (mouse monoclonal, 1:1,000; Sigma), STEM121 (mouse monoclonal, 1:300; StemCells). Anti-mouse or rabbit secondary antibodies conjugated to Alexa Fluor-488 or Alexa Fluor-568 (Life Technologies) were used and counterstained with 49,69-diamidino-2-phenylindole (DAPI). Images were taken with a confocal microscope (Eclipse C1si; Nikon Instruments, Inc., Melville, NY).

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Tsai Y., Lu B., Bakondi B., Girman S., Sahabian A., Sareen D., Svendsen C.N, & Wang S. (2015). Human iPSC-derived neural progenitors preserve vision in an AMD-like model. Stem cells (Dayton, Ohio), 33(8), 2537-2549.

Publication 2015

Cone-arrestin recoverin calbindin D28k parvalbumin synaptophysin RPE65 type IV collagen MAB1281 nestin GFAP Alexa Fluor-488 Alexa Fluor-568 Eclipse C1si

Alexa 568 Alexa fluor 488 Antibodies Arrestin Calbindin d28k Cone Confocal microscope Gfap Mouse Nestin Neurofilament Parvalbumin Pkcα Rabbit Recoverin Retinal S100β Synaptophysin Type iv collagen

Corresponding Organization :

Other organizations : Cedars-Sinai Medical Center

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Variable analysis

independent variables

Antibodies used for staining: cone-arrestin, PKCα, recoverin, calbindin D28k, neurofilament-RT97, parvalbumin, synaptophysin, RPE65, type IV collagen, MAB1281, nestin, GFAP, S100β, Ki67, TuJ1, STEM121

dependent variables

Staining patterns and localization of the various markers in the retinal sections

control variables

Published protocols (34, 35) were used for the staining procedures
Anti-mouse or rabbit secondary antibodies conjugated to Alexa Fluor-488 or Alexa Fluor-568 were used
Counterstaining with DAPI was performed
Images were taken with a confocal microscope (Eclipse C1si; Nikon Instruments, Inc., Melville, NY)

controls

Positive controls: Not explicitly mentioned
Negative controls: Not explicitly mentioned

Annotations

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