rabbit anti-opn4  (ATS Bio)

Journal: Frontiers in Neuroanatomy

Article Title: Alpha retinal ganglion cells in pigmented mice retina: number and distribution

doi: 10.3389/fnana.2022.1054849

Figure Lengend Snippet: Primary and secondary antibodies used in this work.

Article Snippet: , Rabbit anti OPN4 , AB-N39 Advanced Targeting Systems , 1:1,000.

Techniques:

Journal: Frontiers in Neuroanatomy

Article Title: Alpha retinal ganglion cells in pigmented mice retina: number and distribution

doi: 10.3389/fnana.2022.1054849

Figure Lengend Snippet: ONs-αRGCs are not immunodetected against melanopsin. Photomicrographs of flat-mounted retinas (A–A ”’ ) or radial retinal sections (B–B ” ) immunodetected against the αRGC pan-marker OPN (A,B) in blue, Calbindin (A’,B’) in green and OPN4 (A”,B”) in red (merged in A ”’ ) in the same area of the retina. White circles represent cells positive for OPN + and Calbindin + but negative for OPN4 (ONs-αRGCs/M4ipRGCs) and yellow arrowheads represent cells positive for OPN4 but negative for OPN and Calbindin (M1-M3-ipRGCs). Scale bar: 100 μm.

Article Snippet: , Rabbit anti OPN4 , AB-N39 Advanced Targeting Systems , 1:1,000.

Techniques: Marker

Journal: Journal of Cell Science

Article Title: Divergent G-protein selectivity across melanopsins from mice and humans

doi: 10.1242/jcs.258474

Figure Lengend Snippet: Human melanopsin shows light-dependent coupling of both Gα i/o and Gα s . (A) Immunocytochemistry photomicrograph showing human OPN4 (green) in HEK293T cells (DAPI-stained nuclei, blue). (B–F) Changes in bioluminescence in response to a 1 s 470 nm light flash (arrow), normalised to 1 at the time of the light pulse, from HEK293 cells expressing the cAMP reporter GloSensor and either hOPN4 (B,D–F) or hRod Opsin (C). (B–E) Responses across a range of flash intensities for hOPN4 (B) and hRod Opsin (C) in HEK293T cells, hOPN4 in HEK293T cells treated with pertussis toxin (PTX) to eliminate G i/o signalling (D) and hOPN4 in HEK293 ΔGs/Gq/G12 cells (E). Key for light intensity in B–E is shown below B (B and C, n =4; D and E, n =3). (F) hOPN4-expressing HEK293 ΔGs/Gq/G12 cells treated with PTX with (hOPN4+Gs) or without (hOPN4 only) heterologous Gα s exposed to a 1s 14.09 log photon/cm 2 /sec light flash (hOPN4 only, n =3; hOPN4+Gs, n =5). Data expressed as mean±s.e.m. Cells pretreated with 2 µM forskolin to elevate the starting level of cAMP for traces in B,C,E. NLU, normalised luminescence units. n values denote biological replicates from independent transfections.

Article Snippet: Fixed cells were permeabilised in 10% Triton-X100 (Sigma-Aldrich), blocked for 30 min in phosphate-buffered saline (PBS) containing 5% serum (donkey) prior to incubation at room temperature for 1 h with the relevant primary antibody [anti-1D4 mouse IgG (Abcam, AB5417) used at 1:500 for hOPN4, musOpn4S and musOpn4L in A and Fig. S2 ; anti-mouse Opn4 rabbit polyclonal (ATS-Bio, AB-N39) used at 1:1000 for musOpn4S and musOpn4SL in A] in 1% donkey serum, washed in PBS and incubated for a further 30 min with 10 μg/ml Alexa Fluor 488-conjugated secondary antibody (donkey anti-mouse or anti-rabbit IgG; Life Technologies).

Techniques: Immunocytochemistry, Staining, Expressing, Transfection

Journal: Journal of Cell Science

Article Title: Divergent G-protein selectivity across melanopsins from mice and humans

doi: 10.1242/jcs.258474

Figure Lengend Snippet: Gα s signalling in splice variants of mouse melanopsin. (A) Immunocytochemistry photomicrograph showing musOpn4L (green, top) and musOpn4S (green, bottom) in HEK293T cells (DAPI-stained nuclei, blue). (B–I) Changes in bioluminescence in response to a 1s flash of light (arrow), normalised to 1 at the time of stimulus, from HEK293 cells expressing the cAMP reporter GloSensor and either musOpn4L (B,D,F,H) or musOpn4S (C,E,G,I). (B,C) Responses across a range of flash intensities (key below the graphs) for musOpn4L (B, top) and musOpn4S (C, top). n =4. Brackets indicate regions shown beneath with constrained y axes. (D,E) HEK293T cells treated with PTX to eliminate G i/o signalling or (F,G) HEK293 ΔGs/Gq/G12 cells exposed to a 470 nm flash at a range of light intensities (key below B and C). D, n =4; E, n =5; F, n =3; G, n =4. (H,I) HEK293 ΔGs/Gq/G12 cells treated with PTX with (blue in H, green in I) or without (black) transfection of a Gα s expression vector, exposed to a 1 s 470 nm 14.09 log photon/cm 2 /sec light flash (H, n =4; I, n =4). Data expressed as mean±s.e.m. Cells pretreated with 2 µM forskolin to elevate starting level of cAMP for traces in B,C,F,G. NLU, normalised luminescence units. n values denote biological replicates from independent transfections.

Article Snippet: Fixed cells were permeabilised in 10% Triton-X100 (Sigma-Aldrich), blocked for 30 min in phosphate-buffered saline (PBS) containing 5% serum (donkey) prior to incubation at room temperature for 1 h with the relevant primary antibody [anti-1D4 mouse IgG (Abcam, AB5417) used at 1:500 for hOPN4, musOpn4S and musOpn4L in A and Fig. S2 ; anti-mouse Opn4 rabbit polyclonal (ATS-Bio, AB-N39) used at 1:1000 for musOpn4S and musOpn4SL in A] in 1% donkey serum, washed in PBS and incubated for a further 30 min with 10 μg/ml Alexa Fluor 488-conjugated secondary antibody (donkey anti-mouse or anti-rabbit IgG; Life Technologies).

Techniques: Immunocytochemistry, Staining, Expressing, Transfection, Plasmid Preparation

Journal: Neuron

Article Title: Single-cell profiles of retinal ganglion cells differing in resilience to injury reveal neuroprotective genes

doi: 10.1016/j.neuron.2019.11.006

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit polyclonal anti-Opn4 , Thermo Scientific , Cat#PA1–780;RRID:AB_2267547.

Techniques: Recombinant, Multiplex Assay, RNAscope, Over Expression, Cloning, Plasmid Preparation, Software, Microscopy