Retinal Cell Immunodetection Protocol

Microglial cells were detected using a rabbit monoclonal anti-Iba1 antibody (1:1000–500; ab178846: Abcam, Cambridge, UK). Astrocytes and Müller cells were detected with a goat monoclonal anti- Glial fibrillary acidic protein (GFAP) antibody (1:500; 019-19741: Abcam, Cambridge, UK). The synaptic connections were detected with a mouse monoclonal anti-Bassoon antibody (1:750; ADI-VAM-PS003; Enzo life Science, Lausen, Switzerland). The L-cones, S-cones and rods outer segments were detected using a rabbit monoclonal anti-L/M-opsin (1:1200; ab5405; Chemicon-Millipore Iberica, Madrid, Spain), a goat monoclonal anti-S-opsin (1:1000; N-20; anti-OPN1SW; Santa Cruz Biotechnology, Heidelberg, Germany) and a mouse monoclonal anti-rhodopsin (1:1200, 1D4; Sigma-Aldrich, Madrid, Spain) antibody, respectively. Cone photoreceptors were detected using a rabbit monoclonal anti-cone arrestin antibody (1:1000; AB15282, Merck, Germany). Signs of Oxidative stress were detected with a monoclonal antibody against mouse α-8-hydroxy-2′-deoxyguanosine (8-OhdG; 1:1000, sc-66036; Santa Cruz Biotechnology, Heidelberg, Germany), which is a marker that binds oxidatively damaged proteins and DNA [75 (link)]. Retinal ganglion cells were detected using a goat polyclonal anti-Brn3a (1:750; C-20; Santa Cruz Biotechnology).

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Martínez-Vacas A., Di Pierdomenico J., Valiente-Soriano F.J., Vidal-Sanz M., Picaud S., Villegas-Pérez M.P, & García-Ayuso D. (2021). Glial Cell Activation and Oxidative Stress in Retinal Degeneration Induced by β-Alanine Caused Taurine Depletion and Light Exposure. International Journal of Molecular Sciences, 23(1), 346.

Publication 2021

ab178846 ADI-VAM-PS003 AB15282 sc-66036

8 ohdg Anti antibody Anti glial fibrillary acidic protein antibody Antibody Arrestin Astrocytes Cone Goat Microglial cells Monoclonal antibody Mouse Müller cells Opsin Oxidative stress Proteins Rabbit Retinal ganglion cells Rhodopsin Rods outer segments

Corresponding Organization :

Other organizations : Instituto Murciano de Investigación Biosanitaria, Universidad de Murcia, Inserm, Sorbonne Université, Centre National de la Recherche Scientifique, Institut de la Vision

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Variable analysis

independent variables

Antibody used to detect microglial cells (rabbit monoclonal anti-Iba1 antibody)
Antibody used to detect astrocytes and Müller cells (goat monoclonal anti-GFAP antibody)
Antibody used to detect synaptic connections (mouse monoclonal anti-Bassoon antibody)
Antibody used to detect L-cones, S-cones and rods outer segments (rabbit monoclonal anti-L/M-opsin, goat monoclonal anti-S-opsin, and mouse monoclonal anti-rhodopsin antibodies)
Antibody used to detect cone photoreceptors (rabbit monoclonal anti-cone arrestin antibody)
Antibody used to detect signs of oxidative stress (monoclonal antibody against mouse α-8-hydroxy-2′-deoxyguanosine (8-OhdG))
Antibody used to detect retinal ganglion cells (goat polyclonal anti-Brn3a)

dependent variables

Presence and distribution of microglial cells
Presence and distribution of astrocytes and Müller cells
Presence and distribution of synaptic connections
Presence and distribution of L-cones, S-cones and rods outer segments
Presence and distribution of cone photoreceptors
Presence and level of oxidative stress
Presence and distribution of retinal ganglion cells

control variables

Not explicitly mentioned

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

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