cone outer segments abca4 mab  (Santa Cruz Biotechnology)

Journal: Methods in Molecular Biology

Article Title: Retinal Degeneration

doi: 10.1007/978-1-4939-8669-9

Figure Lengend Snippet: Fig. 2 Human (A, B) and mouse (C) retina whole mounts. (A) Fluorescence image of rhodopsin (Rho 1D4) label- ing of rods surrounding unlabeled cones in a human retinal whole mount. Cones are unlabeled and appear as holes surrounded by labeled rods. (B) Fluorescence image of ABCA4 (Rim 3F4) labeling of cones surrounded by labeled rods [10]. (C) Fluorescence image of cone opsin (blue and red/green opsin) labeling of cone photo- receptors in a mouse retinal whole mount

Article Snippet: Cryosections Table 2 Immunochemical markers for the identification of proteins and cellular compartments of mouse rod and cone photoreceptors Photoreceptor cell layer Antigen Antibody Source References Rod outer segment Rhodopsin Mab: Rho 1D4; Rho 4D2 Millipore [12, 13] Rod outer segment CNGA1 Mab: PMc1D1 Millipore [16, 24] Rod outer segment CNGB1 and GARP proteins Mab: Garp 4B1 Millipore [24, 25] Cone outer segments (M/L) Cone opsin Pab: Cone M/L Millipore [16] Rod and cone outer segments RetGC-1 Mab: GC1-8A5 Millipore [16] Rod and cone outer segments ABCA4 Mab: Rim 3F4; Rim 5B4 Millipore; Santa Cruz [12, 16] Rod and cone outer segments Peripherin-2 (peripherin/rds) Mab: Per 5H2 Millipore [16] Cilium/axoneme Retinitis pigmentosa 1 protein (RP1) Chicken anti-RP1 clone EAP15A Liu et al., 2004 [26] Photoreceptor inner segment Sodium/potassium ATPase α3 Mouse anti-Na/K ATPase clone XVIF9-G10 Abcam [10] Endoplasmic reticulum Calnexin Pab MilliporeSigma Endoplasmic reticulum KDEL Mab:10C3 Enzo, Stressgene [16] CNGA1 cyclic nucleotide-gated channel (Subunit A1), RetGC1 guanylate cyclase −1, Mab monoclonal antibody, Pab polyclonal antibody, GARP glutamic acid-rich proteins Retinal Cell Specific Markers for Immunofluorescence Microscopy 300 8.

Techniques: Fluorescence, Labeling

Journal: Methods in Molecular Biology

Article Title: Retinal Degeneration

doi: 10.1007/978-1-4939-8669-9

Figure Lengend Snippet: Fig. 5 Immunofluorescence of mouse retinal cryosections labeled with antibodies to various proteins localized in photoreceptors. (A) Labeling of wild-type (WT) mouse retina section with Rim 5B4 antibody to ABCA4, the ATP-binding cassette transporter found primarily in photoreceptor cells. (B) Labeling of ABCA4 knockout (KO) mouse section with Rim 5B4 antibody as a control. Labeling of outer segments is absent confirming the speci- ficity of the Rim 5B4 to ABCA4. Some autofluorescence is observed in the RPE cells arising from bisretinoid compounds such as A2E which accumulate in RPE cells of ABCA4 KO mice. (C) Labeling of WT mouse section with RetGC1 8A5 against guanylate cyclase 1 present in rod and cone outer segments. (D) Labeling of WT mouse section with calnexin antibody showing the location of the endoplasmic reticulum (ER) in photorecep- tors just above the ONL. Arrows in (A) and (C) show cone OS labeling. Labeling in (A–C) just below the ONL are blood cells with surface Ig labeled with the secondary antibody against mouse Ig. Bar = 20 μm

Article Snippet: Cryosections Table 2 Immunochemical markers for the identification of proteins and cellular compartments of mouse rod and cone photoreceptors Photoreceptor cell layer Antigen Antibody Source References Rod outer segment Rhodopsin Mab: Rho 1D4; Rho 4D2 Millipore [12, 13] Rod outer segment CNGA1 Mab: PMc1D1 Millipore [16, 24] Rod outer segment CNGB1 and GARP proteins Mab: Garp 4B1 Millipore [24, 25] Cone outer segments (M/L) Cone opsin Pab: Cone M/L Millipore [16] Rod and cone outer segments RetGC-1 Mab: GC1-8A5 Millipore [16] Rod and cone outer segments ABCA4 Mab: Rim 3F4; Rim 5B4 Millipore; Santa Cruz [12, 16] Rod and cone outer segments Peripherin-2 (peripherin/rds) Mab: Per 5H2 Millipore [16] Cilium/axoneme Retinitis pigmentosa 1 protein (RP1) Chicken anti-RP1 clone EAP15A Liu et al., 2004 [26] Photoreceptor inner segment Sodium/potassium ATPase α3 Mouse anti-Na/K ATPase clone XVIF9-G10 Abcam [10] Endoplasmic reticulum Calnexin Pab MilliporeSigma Endoplasmic reticulum KDEL Mab:10C3 Enzo, Stressgene [16] CNGA1 cyclic nucleotide-gated channel (Subunit A1), RetGC1 guanylate cyclase −1, Mab monoclonal antibody, Pab polyclonal antibody, GARP glutamic acid-rich proteins Retinal Cell Specific Markers for Immunofluorescence Microscopy 300 8.

Techniques: Immunofluorescence, Labeling, Binding Assay, Knock-Out, Control

Journal: Methods in Molecular Biology

Article Title: Retinal Degeneration

doi: 10.1007/978-1-4939-8669-9

Figure Lengend Snippet: Fig. 2 TPM imaging of the retina and RPE in mouse models of retinal pathologies in vivo. (a) TPM imaging based on endogenous fluorophores. Left, brightly fluorescent granules of retinal condensation products are visible in the retina and RPE of 3.5-month-old Mertk−/−Abca4−/−Rdh8−/− mice, with defective phagocytosis and retinoid cycle pathways [17]. Right, retinosomes outline the contours of RPE cells in 1-year-old albino Rpe65−/− mice, a mouse model of Leber congenital amaurosis. Both images were obtained with 800 nm laser light. (b) Visualization of retinal capillaries in vivo, in 1-year-old C57BL/6J-TyrC-2J mice, after tail vain injection with a fluorescein—BSA solution; this approach can be used to evaluate leakage from retinal capillaries in mice with induced diabetes. Both left and right images are from different sections of the retina in the same eye. Images were obtained with 820 nm laser light

Article Snippet: Cryosections Table 2 Immunochemical markers for the identification of proteins and cellular compartments of mouse rod and cone photoreceptors Photoreceptor cell layer Antigen Antibody Source References Rod outer segment Rhodopsin Mab: Rho 1D4; Rho 4D2 Millipore [12, 13] Rod outer segment CNGA1 Mab: PMc1D1 Millipore [16, 24] Rod outer segment CNGB1 and GARP proteins Mab: Garp 4B1 Millipore [24, 25] Cone outer segments (M/L) Cone opsin Pab: Cone M/L Millipore [16] Rod and cone outer segments RetGC-1 Mab: GC1-8A5 Millipore [16] Rod and cone outer segments ABCA4 Mab: Rim 3F4; Rim 5B4 Millipore; Santa Cruz [12, 16] Rod and cone outer segments Peripherin-2 (peripherin/rds) Mab: Per 5H2 Millipore [16] Cilium/axoneme Retinitis pigmentosa 1 protein (RP1) Chicken anti-RP1 clone EAP15A Liu et al., 2004 [26] Photoreceptor inner segment Sodium/potassium ATPase α3 Mouse anti-Na/K ATPase clone XVIF9-G10 Abcam [10] Endoplasmic reticulum Calnexin Pab MilliporeSigma Endoplasmic reticulum KDEL Mab:10C3 Enzo, Stressgene [16] CNGA1 cyclic nucleotide-gated channel (Subunit A1), RetGC1 guanylate cyclase −1, Mab monoclonal antibody, Pab polyclonal antibody, GARP glutamic acid-rich proteins Retinal Cell Specific Markers for Immunofluorescence Microscopy 300 8.

Techniques: Imaging, In Vivo, Injection

Journal: Methods in Molecular Biology

Article Title: Retinal Degeneration

doi: 10.1007/978-1-4939-8669-9

Figure Lengend Snippet: Fig. 3 Ex vivo TPM imaging of the retina after induction of photoreceptor damage in intact eyes of 1-month-old albino Abca4−/−Rdh8−/− mice. Reconstructed volumes obtained from a series of z-stack images obtained with 730 nm excitation light are shown in both (a) and (b). The RPE is at z = 0, and the section outlined in blue is 10 μm under the RPE (toward the ganglion cell layer). Red arrows indicate enlarged photoreceptors [12], yel- low arrows point to RPE nuclei. (a) Retinal 3D volume from a mouse treated with DMSO before induction of photoreceptor damage. (b) Retinal 3D volume from a mouse treated with an investigative pharmaceutical before induction of photoreceptor damage. Large quantities of enlarged photoreceptors present in mice treated with DMSO indicate severe damage to the retina

Article Snippet: Cryosections Table 2 Immunochemical markers for the identification of proteins and cellular compartments of mouse rod and cone photoreceptors Photoreceptor cell layer Antigen Antibody Source References Rod outer segment Rhodopsin Mab: Rho 1D4; Rho 4D2 Millipore [12, 13] Rod outer segment CNGA1 Mab: PMc1D1 Millipore [16, 24] Rod outer segment CNGB1 and GARP proteins Mab: Garp 4B1 Millipore [24, 25] Cone outer segments (M/L) Cone opsin Pab: Cone M/L Millipore [16] Rod and cone outer segments RetGC-1 Mab: GC1-8A5 Millipore [16] Rod and cone outer segments ABCA4 Mab: Rim 3F4; Rim 5B4 Millipore; Santa Cruz [12, 16] Rod and cone outer segments Peripherin-2 (peripherin/rds) Mab: Per 5H2 Millipore [16] Cilium/axoneme Retinitis pigmentosa 1 protein (RP1) Chicken anti-RP1 clone EAP15A Liu et al., 2004 [26] Photoreceptor inner segment Sodium/potassium ATPase α3 Mouse anti-Na/K ATPase clone XVIF9-G10 Abcam [10] Endoplasmic reticulum Calnexin Pab MilliporeSigma Endoplasmic reticulum KDEL Mab:10C3 Enzo, Stressgene [16] CNGA1 cyclic nucleotide-gated channel (Subunit A1), RetGC1 guanylate cyclase −1, Mab monoclonal antibody, Pab polyclonal antibody, GARP glutamic acid-rich proteins Retinal Cell Specific Markers for Immunofluorescence Microscopy 300 8.

Techniques: Ex Vivo, Imaging