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goat polyclonal anti unc5b (R&D Systems)

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R&D Systems goat polyclonal anti unc5b
Goat Polyclonal Anti Unc5b, supplied by R&D Systems, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/goat+polyclonal+anti+unc5b/UNC5H2%2FUNC5B+Antibody/bio_rxiv__2020__08__06__234906-208-33-36
Average 90 stars, based on 1 article reviews

goat polyclonal anti unc5b - by Bioz Stars, 2026-09

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Article Title: Expression of netrin-1 receptors in retina of oxygen-induced retinopathy in mice
Article Snippet: The primary antibodies included goat polyclonal anti-UNC5B (AF1006 1:50, R&D Systems, MN, USA), goat polyclonal anti-UNC5C (SC-54441 1:100, Santa Cruz Biotechnology, CA, USA), rabbit polyclonal anti-UNC5D (SC-135262 1:100, Santa Cruz Biotechnology), rabbit polyclonal anti-neogenin (SC-15337 1:100, Santa Cruz Biotechnology), rabbit polyclonal anti-DCC (SC-35184 1:100, Santa Cruz Biotechnology), rabbit polyclonal anti-A2b (SC-28996 1:100, Santa Cruz Biotechnology), and mouse anti-GAPDH (Mab-2005079 1:8000, ProMab, CA, USA).

Immunolabeling:

Article Title: Paranode stability requires UNC5B expression by oligodendrocytes
Article Snippet: .. The optic nerve was collected, post-fixed in 30% sucrose and fixed in OCT. 30 μm sections of optic nerve were immunolabeled using the following antibodies: mouse monoclonal anti-Caspr (UCDavis/NIH NeuroMab Facility, CA, USA); goat polyclonal anti-UNC5B (R&D Systems, MN, USA); Alexa 555-conjugated donkey anti-goat; and Alexa 647-conjugated donkey anti-mouse (Invitrogen, CA, USA). .. Images were taken using an SR-350 Vutara microsope and d-STORM imaging buffer: 1M cysteamine, 2-mercaptoethanol, glucose oxidase, catalase, 1M Tris (pH 8.0), sodium chloride, D-glucose.

Incubation:

Article Title: Role of unc5b in retinal neovascularization in mice with oxygen-induced retinopathy
Article Snippet: Proteins were blotted onto a polyvinylidene difluoride microporous membrane (Millipore, USA). .. The membranes were blocked with 5% nonfat milk at 4°C overnight, and then incubated with goat polyclonal anti-unc5b (1:50, R&D system, USA) or rabbit polyclonal anti-GAPDH antibody (1:3000, Santa Cruz Biotech, USA) for 2 hours at room temperature, followed by incubation with a horseradish peroxidase-conjuncted secondary antibody (1:10000, sigma,USA) for 1 hour at room temperature. .. Protein blots were detected using an enhanced chemiluminescence (ECL) kit (Pierce, USA).

Image Search Results

(A) Real-time RT-PCR analysis of unc5b and plp expression in the mouse brain. (B) Western blot of cerebellum lysates reveals readily detectable amounts of UNC5B protein in the adult CNS. (C) In situ hybridization of unc5b mRNA indicates expression in white matter tracts of corpus callosum and cerebellum. PLP expression is shown for comparison (adapted from the Allen Mouse Brain Atlas (Lein et al., 2007); ( http://mouse.brainmap.org/gene/show/18587 and http://mouse.brain-map.org/experiment/show/71656860 ). Scale bar: 420 and 210 μm. (D) UNC5B and MBP immunostaining of mature oligodendrocytes cultured in vitro for 8 days. Scale bar: 32.5 μm. (E) Western blot analysis of UNC5B in cultures of purified oligodendrocytes at 1, 5 and 8 DIV. (F) Caspr1 and UNC5B immunostaining of myelinated axons teased from adult mouse spinal cord, showing UNC5B and Caspr1 proteins enriched at the paranode. Scale bar: 3 μm. (G-I) STORM super-resolution imaging of a section of adult mouse optic nerve revealing UNC5B (green) immunostaining spiraling around the Caspr1 (magenta) positive domains. H and I correspond to the same paranode. Scale bar: G, H and I: 1 μm; J: 700 nm.

Journal: bioRxiv

Article Title: Paranode stability requires UNC5B expression by oligodendrocytes

doi: 10.1101/2020.08.06.234906

Figure Lengend Snippet: (A) Real-time RT-PCR analysis of unc5b and plp expression in the mouse brain. (B) Western blot of cerebellum lysates reveals readily detectable amounts of UNC5B protein in the adult CNS. (C) In situ hybridization of unc5b mRNA indicates expression in white matter tracts of corpus callosum and cerebellum. PLP expression is shown for comparison (adapted from the Allen Mouse Brain Atlas (Lein et al., 2007); ( http://mouse.brainmap.org/gene/show/18587 and http://mouse.brain-map.org/experiment/show/71656860 ). Scale bar: 420 and 210 μm. (D) UNC5B and MBP immunostaining of mature oligodendrocytes cultured in vitro for 8 days. Scale bar: 32.5 μm. (E) Western blot analysis of UNC5B in cultures of purified oligodendrocytes at 1, 5 and 8 DIV. (F) Caspr1 and UNC5B immunostaining of myelinated axons teased from adult mouse spinal cord, showing UNC5B and Caspr1 proteins enriched at the paranode. Scale bar: 3 μm. (G-I) STORM super-resolution imaging of a section of adult mouse optic nerve revealing UNC5B (green) immunostaining spiraling around the Caspr1 (magenta) positive domains. H and I correspond to the same paranode. Scale bar: G, H and I: 1 μm; J: 700 nm.

Article Snippet: The following antibodies were used in this study: mouse monoclonal anti-α-actin (Sigma), mouse monoclonal anti-Caspr1 for immunohistochemistry (UCDavis/NIH NeuroMab Facility), rabbit anti-Kv1.1 (Abcam), mouse anti-pan sodium channel (Sigma), mouse monoclonal anti β-tubulin III (Abcam), goat polyclonal anti-UNC5B for immunostaining (R&D Research), rabbit monoclonal anti-UNC5B for western blot (Cell Signaling), rabbit monoclonal anti-Claudin-11 (Abcam) and rabbit monoclonal anti-JAM-C (Abcam).

Techniques: Quantitative RT-PCR, Expressing, Western Blot, In Situ Hybridization, Immunostaining, Cell Culture, In Vitro, Purification, Imaging

(A) Schematic representation of the floxed unc5b allele and (B) representative image showing genotyping of unc5bFlox mice. (C) β-Gal staining of brain sections of adult Olig2-Cre/ROSA26 reporter mice shows Olig2-driven Cre expression in oligodendrocytes of corpus callosum and cerebellum. (D) RT-PCR analysis of unc5b expression in brain stem lysates of 3 month-old cKO mice shows reduced unc5b mRNA expression compared to WT controls. (E) Western blot analysis of 3 and 12 month-old mouse optic nerve, spinal cord and brain stem lysates confirms deletion of unc5b expression from oligodendrocytes.

Journal: bioRxiv

Article Title: Paranode stability requires UNC5B expression by oligodendrocytes

doi: 10.1101/2020.08.06.234906

Figure Lengend Snippet: (A) Schematic representation of the floxed unc5b allele and (B) representative image showing genotyping of unc5bFlox mice. (C) β-Gal staining of brain sections of adult Olig2-Cre/ROSA26 reporter mice shows Olig2-driven Cre expression in oligodendrocytes of corpus callosum and cerebellum. (D) RT-PCR analysis of unc5b expression in brain stem lysates of 3 month-old cKO mice shows reduced unc5b mRNA expression compared to WT controls. (E) Western blot analysis of 3 and 12 month-old mouse optic nerve, spinal cord and brain stem lysates confirms deletion of unc5b expression from oligodendrocytes.

Techniques: Staining, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot

(A) Western blot analysis of major myelin proteins PLP and MBP in optic nerve, spinal cord and brain stem lysates of 6-9 month old cKO and wild-type littermates. (B,C) Percent of myelinated axons and (D,E) g-ratios were measured in electron micrographs of coronal sections of optic nerve from 7 month old UNC5B cKO and wild-type mice. Scale bar: (B) – 2 μm (D) – 300 nm. 450-650 axons were analyzed to calculate percent of myelinated axons; 40-50 axons were analyzed for g-ratio quantification. (F) Representative micrographs of wild-type and cKO optic nerve sections showing general features of compact myelin. Scale bar: 40 nm (G) Mean periodicity is reduced by deletion of unc5b expression. >100 individual periods were measured for both genotypes. (H, I) Intensity profiles across the line at the center of the micrograph (spanning 2 periods) were collected for 20-25 micrographs and averaged. The vertical dotted lines represent the end of a myelin period. Myelin periodicity is ~1 nm shorter in UNC5B cKOs. Scale bar in (H) : 10 nm. (J) The number of myelin outfoldings and the area of the inner tongue in EM cross-sections of the optic nerve are comparable in UNC5B cKO and wild-type animals. Inner tongue area was analyzed in 20 axons. Bar graphs are plots of means and error bars indicate ±SEM. **** p < 0.0001 (unpaired student-t test).

Journal: bioRxiv

Article Title: Paranode stability requires UNC5B expression by oligodendrocytes

doi: 10.1101/2020.08.06.234906

Figure Lengend Snippet: (A) Western blot analysis of major myelin proteins PLP and MBP in optic nerve, spinal cord and brain stem lysates of 6-9 month old cKO and wild-type littermates. (B,C) Percent of myelinated axons and (D,E) g-ratios were measured in electron micrographs of coronal sections of optic nerve from 7 month old UNC5B cKO and wild-type mice. Scale bar: (B) – 2 μm (D) – 300 nm. 450-650 axons were analyzed to calculate percent of myelinated axons; 40-50 axons were analyzed for g-ratio quantification. (F) Representative micrographs of wild-type and cKO optic nerve sections showing general features of compact myelin. Scale bar: 40 nm (G) Mean periodicity is reduced by deletion of unc5b expression. >100 individual periods were measured for both genotypes. (H, I) Intensity profiles across the line at the center of the micrograph (spanning 2 periods) were collected for 20-25 micrographs and averaged. The vertical dotted lines represent the end of a myelin period. Myelin periodicity is ~1 nm shorter in UNC5B cKOs. Scale bar in (H) : 10 nm. (J) The number of myelin outfoldings and the area of the inner tongue in EM cross-sections of the optic nerve are comparable in UNC5B cKO and wild-type animals. Inner tongue area was analyzed in 20 axons. Bar graphs are plots of means and error bars indicate ±SEM. **** p < 0.0001 (unpaired student-t test).

Techniques: Western Blot, Expressing

(A) Electron micrographs of longitudinal sections of 7 month-old wild-type and UNC5B cKO optic nerves. Top left panel shows a micrograph of a wild-type paranode and bottom two panels show examples of highly disorganized UNC5B cKO paranodes. The boxed area in the left panel is magnified on the right. Note the segmented loop-loop interface (arrowheads) and large vesicles (asterisks) inside glial loops of cKO paranodes. Scale bars: 100 nm. (B) Mean number of detached loops per paranode. ** p < 0.01 (Mann-Whitney test). 31 WT and 33 UNC5B cKO paranodes were analyzed. (C) Paranode distribution across different levels of disorganization. (D) Mean number of gaps detected on glial loop-loop interfaces. * p < 0.05 (Mann-Whitney test). 14 wild-type and 19 UNC5B cKO paranodes were analyzed. Bar graphs are plots of means and error bars indicate ±SEM. (E) 200 nm longitudinal sections of optic nerve were imaged under a TEM. The microscope stage was tilted by 2-5 degrees between images; tilting progressively reveals fragmentation (white arrowheads) of the glial loop interface (upper panels), which invades the compact myelin (bottom panels). The arrows indicate the direction of tilting of the stage. Scale bar: 100 nm.

Journal: bioRxiv

Article Title: Paranode stability requires UNC5B expression by oligodendrocytes

doi: 10.1101/2020.08.06.234906

Figure Lengend Snippet: (A) Electron micrographs of longitudinal sections of 7 month-old wild-type and UNC5B cKO optic nerves. Top left panel shows a micrograph of a wild-type paranode and bottom two panels show examples of highly disorganized UNC5B cKO paranodes. The boxed area in the left panel is magnified on the right. Note the segmented loop-loop interface (arrowheads) and large vesicles (asterisks) inside glial loops of cKO paranodes. Scale bars: 100 nm. (B) Mean number of detached loops per paranode. ** p < 0.01 (Mann-Whitney test). 31 WT and 33 UNC5B cKO paranodes were analyzed. (C) Paranode distribution across different levels of disorganization. (D) Mean number of gaps detected on glial loop-loop interfaces. * p < 0.05 (Mann-Whitney test). 14 wild-type and 19 UNC5B cKO paranodes were analyzed. Bar graphs are plots of means and error bars indicate ±SEM. (E) 200 nm longitudinal sections of optic nerve were imaged under a TEM. The microscope stage was tilted by 2-5 degrees between images; tilting progressively reveals fragmentation (white arrowheads) of the glial loop interface (upper panels), which invades the compact myelin (bottom panels). The arrows indicate the direction of tilting of the stage. Scale bar: 100 nm.

Techniques: MANN-WHITNEY, Microscopy

Measurement of the length of immunoreactive domains and extent of overlap of Caspr1, Kv1.1, and Na + channel domains along axons in sections of cerebellum and optic nerve of 6-9 month old adult UNC5B cKO and wild-type control mice. (A) Schematic representation of the measurements. (B) Representative images of Caspr1, Kv1.1 and Na + channel staining in the optic nerve of UNC5B cKO and wild-type mice. (C) Caspr1/Na + channel staining of optic nerve showing highly disorganized paranodes in UNC5B cKOs. Arrowheads indicate nodes of Ranvier invaded by Caspr1. (D) Western blot analysis of protein lysates from wild-type and UNC5B cKOs reveals unchanged Caspr expression following Unc5b deletion in oligodendrocytes. (E, F) Length measurements of immunoreactive domains and extent of overlap between adjacent domains in cerebellum and optic nerve. ** p < 0.01; **** p < 0.0001 (unpaired student-t test). 150-200 measurements were taken for each immunoreactive domain. Bar graphs are plots of means and error bars indicate ±SEM.

Journal: bioRxiv

Article Title: Paranode stability requires UNC5B expression by oligodendrocytes

doi: 10.1101/2020.08.06.234906

Figure Lengend Snippet: Measurement of the length of immunoreactive domains and extent of overlap of Caspr1, Kv1.1, and Na + channel domains along axons in sections of cerebellum and optic nerve of 6-9 month old adult UNC5B cKO and wild-type control mice. (A) Schematic representation of the measurements. (B) Representative images of Caspr1, Kv1.1 and Na + channel staining in the optic nerve of UNC5B cKO and wild-type mice. (C) Caspr1/Na + channel staining of optic nerve showing highly disorganized paranodes in UNC5B cKOs. Arrowheads indicate nodes of Ranvier invaded by Caspr1. (D) Western blot analysis of protein lysates from wild-type and UNC5B cKOs reveals unchanged Caspr expression following Unc5b deletion in oligodendrocytes. (E, F) Length measurements of immunoreactive domains and extent of overlap between adjacent domains in cerebellum and optic nerve. ** p < 0.01; **** p < 0.0001 (unpaired student-t test). 150-200 measurements were taken for each immunoreactive domain. Bar graphs are plots of means and error bars indicate ±SEM.

Techniques: Staining, Western Blot, Expressing

(A) MBP and Caspr1 immunostaining in cortical sections from UNC5B cKO and wild-type mice. White arrowheads indicate the end of an internode. Scale bar: 10 μm (B) Internode length quantification. ** p < 0.001 (Mann-Whitney test) 80 internodes per genotype were measured. Bar graphs are plots of means and error bars indicate ±SEM.

Journal: bioRxiv

Article Title: Paranode stability requires UNC5B expression by oligodendrocytes

doi: 10.1101/2020.08.06.234906

Figure Lengend Snippet: (A) MBP and Caspr1 immunostaining in cortical sections from UNC5B cKO and wild-type mice. White arrowheads indicate the end of an internode. Scale bar: 10 μm (B) Internode length quantification. ** p < 0.001 (Mann-Whitney test) 80 internodes per genotype were measured. Bar graphs are plots of means and error bars indicate ±SEM.

Techniques: Immunostaining, MANN-WHITNEY

(A) Representative EM micrographs of optic nerves from 3 month-old UNC5B cKO and wild-type mice. Scale bar: 100 nm. Histograms show (B) mean number of detached loops per paranode and (C) proportion of paranodes with different levels of disorganization. * p < 0.05 (Mann-Whitney test). 50-58 paranodes from 3 animals were analyzed per genotype. (D) Representative images of Caspr1 and Na + channel staining in the cerebellum of 3 month-old UNC5B cKO and wild-type mice. Scale bar: 1 μm. (E, F) Caspr domain length was measured in the cerebellum and cortex. **** p < 0.0001 (unpaired Student’s t-test). (G) MBP and Caspr staining on cortical sections from UNC5B cKO and wild-type mice. White arrowheads indicate internode endings. Scale bar: 10 μm. (H) MBP domain length. 80 internodes per genotype were measured. (I) Myelin periodicity was measured in 32 and 28 axons (~150 periods) from WT and UNC5B cKO animals, respectively. Bar graphs are plots of means and error bars indicate ±SEM.

Journal: bioRxiv

Article Title: Paranode stability requires UNC5B expression by oligodendrocytes

doi: 10.1101/2020.08.06.234906

Figure Lengend Snippet: (A) Representative EM micrographs of optic nerves from 3 month-old UNC5B cKO and wild-type mice. Scale bar: 100 nm. Histograms show (B) mean number of detached loops per paranode and (C) proportion of paranodes with different levels of disorganization. * p < 0.05 (Mann-Whitney test). 50-58 paranodes from 3 animals were analyzed per genotype. (D) Representative images of Caspr1 and Na + channel staining in the cerebellum of 3 month-old UNC5B cKO and wild-type mice. Scale bar: 1 μm. (E, F) Caspr domain length was measured in the cerebellum and cortex. **** p < 0.0001 (unpaired Student’s t-test). (G) MBP and Caspr staining on cortical sections from UNC5B cKO and wild-type mice. White arrowheads indicate internode endings. Scale bar: 10 μm. (H) MBP domain length. 80 internodes per genotype were measured. (I) Myelin periodicity was measured in 32 and 28 axons (~150 periods) from WT and UNC5B cKO animals, respectively. Bar graphs are plots of means and error bars indicate ±SEM.

Techniques: MANN-WHITNEY, Staining

(A) Open field analysis of the spontaneous locomotor activity of UNC5B cKO and wild-type littermates. Histograms show (B) distance travelled and (C) mean active time. n =14 WTs and 13 cKOs at 3 months; 14 WT and 8 cKOs at 6-9 months; 8 WTs and 12 cKOs at 12 months. (D, E) Accelerating Rotarod test. Histograms show (D) total number of falls on each day of training and (E) time mice spent on the rotarod before falling on the test day. n = 13 WTs and 13 cKOs at 3 months; 15 WTs and 8 cKOs at 6-9 months; 12 WTs and 17 cKOs at 12 months. * p < 0.05 (Repeated measures 2 way-ANOVA, followed by Sidak’s multiple comparison test). Bar graphs are plots of means and error bars indicate ±SEM.

Journal: bioRxiv

Article Title: Paranode stability requires UNC5B expression by oligodendrocytes

doi: 10.1101/2020.08.06.234906

Figure Lengend Snippet: (A) Open field analysis of the spontaneous locomotor activity of UNC5B cKO and wild-type littermates. Histograms show (B) distance travelled and (C) mean active time. n =14 WTs and 13 cKOs at 3 months; 14 WT and 8 cKOs at 6-9 months; 8 WTs and 12 cKOs at 12 months. (D, E) Accelerating Rotarod test. Histograms show (D) total number of falls on each day of training and (E) time mice spent on the rotarod before falling on the test day. n = 13 WTs and 13 cKOs at 3 months; 15 WTs and 8 cKOs at 6-9 months; 12 WTs and 17 cKOs at 12 months. * p < 0.05 (Repeated measures 2 way-ANOVA, followed by Sidak’s multiple comparison test). Bar graphs are plots of means and error bars indicate ±SEM.

Techniques: Activity Assay

(A) Representative images of Claudin-11 and Caspr1 immunostaining in optic nerve sections from 6-9 month old UNC5B cKO and age matched wild-type mice. Scale bar: 5 um (B) Quantification of Claudin-11 and Caspr1 fluorescence intensity within the ROI delimited by Caspr1 staining. >300 paranodes were quantified per genotype. **** p < 0.0001 (unpaired Student’s t test) (C,D) Western blot analysis of UNC5B cKO and wild-type optic nerve lysates. Graphs show densitometry analysis of JAM-C and Claudin-11 blots. N= 4 animals per genotype. * p < 0.05, ** p < 0.01 (unpaired Student’s t test). Bar graphs are plots of means and error bars indicate ±SEM.

Journal: bioRxiv

Article Title: Paranode stability requires UNC5B expression by oligodendrocytes

doi: 10.1101/2020.08.06.234906

Figure Lengend Snippet: (A) Representative images of Claudin-11 and Caspr1 immunostaining in optic nerve sections from 6-9 month old UNC5B cKO and age matched wild-type mice. Scale bar: 5 um (B) Quantification of Claudin-11 and Caspr1 fluorescence intensity within the ROI delimited by Caspr1 staining. >300 paranodes were quantified per genotype. **** p < 0.0001 (unpaired Student’s t test) (C,D) Western blot analysis of UNC5B cKO and wild-type optic nerve lysates. Graphs show densitometry analysis of JAM-C and Claudin-11 blots. N= 4 animals per genotype. * p < 0.05, ** p < 0.01 (unpaired Student’s t test). Bar graphs are plots of means and error bars indicate ±SEM.

Techniques: Immunostaining, Fluorescence, Staining, Western Blot

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goat polyclonal anti unc5b (R&D Systems)

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