rabbit anti nt3  (Alomone Labs)


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    Alomone Labs rabbit anti nt3
    Sortilin-mediated binding and internalisation of <t>NT3</t> and proNT3 (A–J) HEK293 cells transfected with Sortilin, p75 NTR or proSortilin as indicated were incubated with 50nM of the given ligand (NT3 (A–C) , proNT3 (D–F) , or GST-NT3pro (G–J) ) for 40 minutes at 37° C. Internalised ligand was visualised by immunofluorescence labelling with specific antibodies. Mature NT3 and proNT3 were detected with anti-NT3, whereas GST-NT3pro was detected with an anti-GST antibody. Cells transfected with Sortilin showed no binding and uptake of NT3 (A) , but internalisation was detected of both proNT3 (D) and GST-NT3pro (G) . The uptake of proNT3 could be inhibited by addition of NTS (E) . Cells transfected with p75 NTR displayed strong surface labelling after incubation with NT3 (C) and proNT3 (F) , but no binding of GST-NT3pro (I) . HEK293 human embryonic kidney 293 cells , NT3 neurotrophin-3 , proNT3 proform of NT3 , p75 NTR p75 neurotrophin receptor , NTS neurotensin , GST-NT3pro prodomain of NT3 coupled to glutathione-s-transferase.
    Rabbit Anti Nt3, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti nt3/product/Alomone Labs
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti nt3 - by Bioz Stars, 2022-10
    92/100 stars

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    1) Product Images from "Proneurotrophin-3 may induce Sortilin dependent death in inner ear neurons"

    Article Title: Proneurotrophin-3 may induce Sortilin dependent death in inner ear neurons

    Journal: The European journal of neuroscience

    doi: 10.1111/j.1460-9568.2010.07556.x

    Sortilin-mediated binding and internalisation of NT3 and proNT3 (A–J) HEK293 cells transfected with Sortilin, p75 NTR or proSortilin as indicated were incubated with 50nM of the given ligand (NT3 (A–C) , proNT3 (D–F) , or GST-NT3pro (G–J) ) for 40 minutes at 37° C. Internalised ligand was visualised by immunofluorescence labelling with specific antibodies. Mature NT3 and proNT3 were detected with anti-NT3, whereas GST-NT3pro was detected with an anti-GST antibody. Cells transfected with Sortilin showed no binding and uptake of NT3 (A) , but internalisation was detected of both proNT3 (D) and GST-NT3pro (G) . The uptake of proNT3 could be inhibited by addition of NTS (E) . Cells transfected with p75 NTR displayed strong surface labelling after incubation with NT3 (C) and proNT3 (F) , but no binding of GST-NT3pro (I) . HEK293 human embryonic kidney 293 cells , NT3 neurotrophin-3 , proNT3 proform of NT3 , p75 NTR p75 neurotrophin receptor , NTS neurotensin , GST-NT3pro prodomain of NT3 coupled to glutathione-s-transferase.
    Figure Legend Snippet: Sortilin-mediated binding and internalisation of NT3 and proNT3 (A–J) HEK293 cells transfected with Sortilin, p75 NTR or proSortilin as indicated were incubated with 50nM of the given ligand (NT3 (A–C) , proNT3 (D–F) , or GST-NT3pro (G–J) ) for 40 minutes at 37° C. Internalised ligand was visualised by immunofluorescence labelling with specific antibodies. Mature NT3 and proNT3 were detected with anti-NT3, whereas GST-NT3pro was detected with an anti-GST antibody. Cells transfected with Sortilin showed no binding and uptake of NT3 (A) , but internalisation was detected of both proNT3 (D) and GST-NT3pro (G) . The uptake of proNT3 could be inhibited by addition of NTS (E) . Cells transfected with p75 NTR displayed strong surface labelling after incubation with NT3 (C) and proNT3 (F) , but no binding of GST-NT3pro (I) . HEK293 human embryonic kidney 293 cells , NT3 neurotrophin-3 , proNT3 proform of NT3 , p75 NTR p75 neurotrophin receptor , NTS neurotensin , GST-NT3pro prodomain of NT3 coupled to glutathione-s-transferase.

    Techniques Used: Binding Assay, Transfection, Incubation, Immunofluorescence

    Crosslinking and precipitation of p75 NTR :Sortilin:proNT3 complexes HEK293 cells transfected with an endocytosis-impaired variant of Sortilin, Sortilin-mut, and p75 NTR as indicated were incubated with or without 50 nM ligand (proNT3 or proNGF) before treatment with crosslinker. Receptor:ligand complexes were immunoprecipitated with anti-p75 NTR , and components were visualised after reducing SDS-PAGE by immunoblotting with anti-Sortilin, anti-proNT3, and anti-p75 NTR . The expression of Sortilin and p75 NTR in input cell lysates were confirmed by immunoblotting. HEK293 human embryonic kidney 293 cells , p75 NTR p75 neurotrophin receptor , Sortilin-mut endocytosis-impaired variant of Sortilin, proNGF proform of nerve growth factor, proNT3 proform of neurotrophin-3, NT3 mature form of neurotrophin-3
    Figure Legend Snippet: Crosslinking and precipitation of p75 NTR :Sortilin:proNT3 complexes HEK293 cells transfected with an endocytosis-impaired variant of Sortilin, Sortilin-mut, and p75 NTR as indicated were incubated with or without 50 nM ligand (proNT3 or proNGF) before treatment with crosslinker. Receptor:ligand complexes were immunoprecipitated with anti-p75 NTR , and components were visualised after reducing SDS-PAGE by immunoblotting with anti-Sortilin, anti-proNT3, and anti-p75 NTR . The expression of Sortilin and p75 NTR in input cell lysates were confirmed by immunoblotting. HEK293 human embryonic kidney 293 cells , p75 NTR p75 neurotrophin receptor , Sortilin-mut endocytosis-impaired variant of Sortilin, proNGF proform of nerve growth factor, proNT3 proform of neurotrophin-3, NT3 mature form of neurotrophin-3

    Techniques Used: Transfection, Variant Assay, Incubation, Immunoprecipitation, SDS Page, Expressing

    Immunoblotting of inner ear tissue for Sortilin, p75 NTR , proNT3 and mature NT3 Subfractions of membranous labyrinth from dissected newborn rats (P3–P5) were pooled, homogenised and centrifuged. The supernatants were subjected to SDS-PAGE followed by immunoblotting for Sortilin, p75 NTR , proNT3 and mature NT3. Beta-actin (~42 kDa) blots are included for evaluation of total protein concentration. (A) Sortilin (~98 kDa) was detected in whole-organ cochleae (Co) and in all in the subfractions i.e. the stria vascularis (I), the organ of Corti with Reissner’s membrane attached (II), the modiolus (not including the spiral ganglion) (III), and the spiral ganglion (IV). (B) P75 NTR (~64 kDa) was detected in the modiolus (III) and the SG (IV). (C) ProNT3 and NT3 were detected in all subfractions. (D) The specificity of proNT3 and NT3 antibodies was tested by immunoblotting of recombinant human proNGF (lane 1 (5 ng) and 2 (1 ng)), proNT3 (lane 3 (5 ng) and 4 (1 ng)), and NT3 (lane 5 (5 ng) and 6 (1 ng)). The proNT3 antibody recognises only proNT3, while the NT3 antibody detects both the proform and the mature form of NT3. kDa kilodalton, Co whole-organ membranous cochlea, p75 NTR p75 neurotrophin receptor, proNT3 proform of neurotrophin-3, NT3 mature form of neurotrophin-3, αSortilin anti-Sortilin, αp75 NTR anti- p75 NTR , αproNT3 anti-proNT3, αNT3 anti-NT3 , αbeta-actin anti-beta actin, P3–P5 postnatal day 3–5
    Figure Legend Snippet: Immunoblotting of inner ear tissue for Sortilin, p75 NTR , proNT3 and mature NT3 Subfractions of membranous labyrinth from dissected newborn rats (P3–P5) were pooled, homogenised and centrifuged. The supernatants were subjected to SDS-PAGE followed by immunoblotting for Sortilin, p75 NTR , proNT3 and mature NT3. Beta-actin (~42 kDa) blots are included for evaluation of total protein concentration. (A) Sortilin (~98 kDa) was detected in whole-organ cochleae (Co) and in all in the subfractions i.e. the stria vascularis (I), the organ of Corti with Reissner’s membrane attached (II), the modiolus (not including the spiral ganglion) (III), and the spiral ganglion (IV). (B) P75 NTR (~64 kDa) was detected in the modiolus (III) and the SG (IV). (C) ProNT3 and NT3 were detected in all subfractions. (D) The specificity of proNT3 and NT3 antibodies was tested by immunoblotting of recombinant human proNGF (lane 1 (5 ng) and 2 (1 ng)), proNT3 (lane 3 (5 ng) and 4 (1 ng)), and NT3 (lane 5 (5 ng) and 6 (1 ng)). The proNT3 antibody recognises only proNT3, while the NT3 antibody detects both the proform and the mature form of NT3. kDa kilodalton, Co whole-organ membranous cochlea, p75 NTR p75 neurotrophin receptor, proNT3 proform of neurotrophin-3, NT3 mature form of neurotrophin-3, αSortilin anti-Sortilin, αp75 NTR anti- p75 NTR , αproNT3 anti-proNT3, αNT3 anti-NT3 , αbeta-actin anti-beta actin, P3–P5 postnatal day 3–5

    Techniques Used: SDS Page, Protein Concentration, Recombinant

    ProNT3 induced apoptosis in SCG neurons The apoptotic potential of proNT3 was tested in SCG neurons. Here is shown the number of apoptotic neurons in percent of the total number counted. SCG neurons were incubated in growth media containing NGF (20ng/ml), NT3 (2ng/ml), proNGF (2ng/ml) or proNT3 (4ng/ml). After 36 hours, SCG cultures were fixed, immunostained with α-Tuj1, α-Sortilin and DAPI and subjected to apoptosis analysis by morphological evaluation. The results demonstrate that NGF (as opposed to NT3) is imperative for SCG neuron survival. In contrast, both proNGF and proNT3 significantly increased the proportion of apoptotic neurons compared to NGF withdrawal alone. SCG superior cervical ganglion, proNT proneurotrophins, NGF nerve growth factor, proNGF proform of NGF , None no (pro)neurotrophins added , NT3 neurotrophin-3, proNT3 proform of NT3.
    Figure Legend Snippet: ProNT3 induced apoptosis in SCG neurons The apoptotic potential of proNT3 was tested in SCG neurons. Here is shown the number of apoptotic neurons in percent of the total number counted. SCG neurons were incubated in growth media containing NGF (20ng/ml), NT3 (2ng/ml), proNGF (2ng/ml) or proNT3 (4ng/ml). After 36 hours, SCG cultures were fixed, immunostained with α-Tuj1, α-Sortilin and DAPI and subjected to apoptosis analysis by morphological evaluation. The results demonstrate that NGF (as opposed to NT3) is imperative for SCG neuron survival. In contrast, both proNGF and proNT3 significantly increased the proportion of apoptotic neurons compared to NGF withdrawal alone. SCG superior cervical ganglion, proNT proneurotrophins, NGF nerve growth factor, proNGF proform of NGF , None no (pro)neurotrophins added , NT3 neurotrophin-3, proNT3 proform of NT3.

    Techniques Used: Incubation

    SPR analysis of ligand binding to immobilised Sortilin SPR analysis of receptor-ligand interactions between the NT3 and immobilised Sortilin demonstrated that mature NT3 binds to Sortilin with a Kd ~20 μM (A) . This affinity is significantly lower than the affinity found for the interaction between proNT3 and Sortilin (Kd ~20 nM) (B) , and for the interaction between the receptor and the prodomain sequence of proNT3 fused to GST (Kd~20 nM) (D) . The interaction between Sortilin and proNT3 can be inhibited by the well-known Sortilin ligand neurotensin (C) . NT3 neurotrophin-3, proNT3 proform of NT3, GST-NT3pro prodomain of NT3 coupled to glutathione-s-transferase NTS neurotensin, Kd dissociation constant, SPR Surface plasmon resonance
    Figure Legend Snippet: SPR analysis of ligand binding to immobilised Sortilin SPR analysis of receptor-ligand interactions between the NT3 and immobilised Sortilin demonstrated that mature NT3 binds to Sortilin with a Kd ~20 μM (A) . This affinity is significantly lower than the affinity found for the interaction between proNT3 and Sortilin (Kd ~20 nM) (B) , and for the interaction between the receptor and the prodomain sequence of proNT3 fused to GST (Kd~20 nM) (D) . The interaction between Sortilin and proNT3 can be inhibited by the well-known Sortilin ligand neurotensin (C) . NT3 neurotrophin-3, proNT3 proform of NT3, GST-NT3pro prodomain of NT3 coupled to glutathione-s-transferase NTS neurotensin, Kd dissociation constant, SPR Surface plasmon resonance

    Techniques Used: SPR Assay, Ligand Binding Assay, Sequencing

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    Alomone Labs rabbit anti nt3
    Sortilin-mediated binding and internalisation of <t>NT3</t> and proNT3 (A–J) HEK293 cells transfected with Sortilin, p75 NTR or proSortilin as indicated were incubated with 50nM of the given ligand (NT3 (A–C) , proNT3 (D–F) , or GST-NT3pro (G–J) ) for 40 minutes at 37° C. Internalised ligand was visualised by immunofluorescence labelling with specific antibodies. Mature NT3 and proNT3 were detected with anti-NT3, whereas GST-NT3pro was detected with an anti-GST antibody. Cells transfected with Sortilin showed no binding and uptake of NT3 (A) , but internalisation was detected of both proNT3 (D) and GST-NT3pro (G) . The uptake of proNT3 could be inhibited by addition of NTS (E) . Cells transfected with p75 NTR displayed strong surface labelling after incubation with NT3 (C) and proNT3 (F) , but no binding of GST-NT3pro (I) . HEK293 human embryonic kidney 293 cells , NT3 neurotrophin-3 , proNT3 proform of NT3 , p75 NTR p75 neurotrophin receptor , NTS neurotensin , GST-NT3pro prodomain of NT3 coupled to glutathione-s-transferase.
    Rabbit Anti Nt3, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti nt3/product/Alomone Labs
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti nt3 - by Bioz Stars, 2022-10
    92/100 stars
      Buy from Supplier

    93
    Alomone Labs nt3
    Inhibition of microglial activation after intravitreal injection of recombinant mature <t>NT3</t> and blockage of P75 NTR . Transgenic mice received twice intravitreal injections of NT3 (0.4 μg), P75 NTR rabbit polyclonal a polyclonal Ab to P75 NTR (1:1 dilution), a combination of both or balanced salt solution ( BSS ) 3d and 6d after tamoxifen (TMX)-induced Muller cell ablation. Eyes were enucleated 10d after TMX treatment for retinal flat-mount staining using an Ab against ionized calcium binding adaptor molecule 1 ( IBA-1 ) for microglia ( A, D, G and J , green ) and peanut-agglutinin ( PNA )-conjugated with Alexa Fluor-594 for cone photoreceptor outer segments ( B, E, H and K , red ). (C, F, I and L) Merged images. (M) Quantitative analysis of IBA-1 stained retinal whole mounts shows intravitreal injection of NT3 and P75 NTR blockage, either alone or in combination, protected photoreceptors concurrently with inhibition of microglial activation. * P
    Nt3, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nt3/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    nt3 - by Bioz Stars, 2022-10
    93/100 stars
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    Sortilin-mediated binding and internalisation of NT3 and proNT3 (A–J) HEK293 cells transfected with Sortilin, p75 NTR or proSortilin as indicated were incubated with 50nM of the given ligand (NT3 (A–C) , proNT3 (D–F) , or GST-NT3pro (G–J) ) for 40 minutes at 37° C. Internalised ligand was visualised by immunofluorescence labelling with specific antibodies. Mature NT3 and proNT3 were detected with anti-NT3, whereas GST-NT3pro was detected with an anti-GST antibody. Cells transfected with Sortilin showed no binding and uptake of NT3 (A) , but internalisation was detected of both proNT3 (D) and GST-NT3pro (G) . The uptake of proNT3 could be inhibited by addition of NTS (E) . Cells transfected with p75 NTR displayed strong surface labelling after incubation with NT3 (C) and proNT3 (F) , but no binding of GST-NT3pro (I) . HEK293 human embryonic kidney 293 cells , NT3 neurotrophin-3 , proNT3 proform of NT3 , p75 NTR p75 neurotrophin receptor , NTS neurotensin , GST-NT3pro prodomain of NT3 coupled to glutathione-s-transferase.

    Journal: The European journal of neuroscience

    Article Title: Proneurotrophin-3 may induce Sortilin dependent death in inner ear neurons

    doi: 10.1111/j.1460-9568.2010.07556.x

    Figure Lengend Snippet: Sortilin-mediated binding and internalisation of NT3 and proNT3 (A–J) HEK293 cells transfected with Sortilin, p75 NTR or proSortilin as indicated were incubated with 50nM of the given ligand (NT3 (A–C) , proNT3 (D–F) , or GST-NT3pro (G–J) ) for 40 minutes at 37° C. Internalised ligand was visualised by immunofluorescence labelling with specific antibodies. Mature NT3 and proNT3 were detected with anti-NT3, whereas GST-NT3pro was detected with an anti-GST antibody. Cells transfected with Sortilin showed no binding and uptake of NT3 (A) , but internalisation was detected of both proNT3 (D) and GST-NT3pro (G) . The uptake of proNT3 could be inhibited by addition of NTS (E) . Cells transfected with p75 NTR displayed strong surface labelling after incubation with NT3 (C) and proNT3 (F) , but no binding of GST-NT3pro (I) . HEK293 human embryonic kidney 293 cells , NT3 neurotrophin-3 , proNT3 proform of NT3 , p75 NTR p75 neurotrophin receptor , NTS neurotensin , GST-NT3pro prodomain of NT3 coupled to glutathione-s-transferase.

    Article Snippet: The blots were blocked in 5% defatted milk in TBST buffer (50mM Tris, 500mM NaCl, 0.1% Tween-20, pH 7.4) for 1 hour at RT and subsequently incubated over night at 4°C in blocking buffer with mouse anti-human Sortilin, rabbit anti-human p75NTR (provided by Moses V. Chao), rabbit anti-proNT3, rabbit anti-NT3 (Alomone Labs Ltd.) or mouse anti-beta-actin (Sigma Aldrich).

    Techniques: Binding Assay, Transfection, Incubation, Immunofluorescence

    Crosslinking and precipitation of p75 NTR :Sortilin:proNT3 complexes HEK293 cells transfected with an endocytosis-impaired variant of Sortilin, Sortilin-mut, and p75 NTR as indicated were incubated with or without 50 nM ligand (proNT3 or proNGF) before treatment with crosslinker. Receptor:ligand complexes were immunoprecipitated with anti-p75 NTR , and components were visualised after reducing SDS-PAGE by immunoblotting with anti-Sortilin, anti-proNT3, and anti-p75 NTR . The expression of Sortilin and p75 NTR in input cell lysates were confirmed by immunoblotting. HEK293 human embryonic kidney 293 cells , p75 NTR p75 neurotrophin receptor , Sortilin-mut endocytosis-impaired variant of Sortilin, proNGF proform of nerve growth factor, proNT3 proform of neurotrophin-3, NT3 mature form of neurotrophin-3

    Journal: The European journal of neuroscience

    Article Title: Proneurotrophin-3 may induce Sortilin dependent death in inner ear neurons

    doi: 10.1111/j.1460-9568.2010.07556.x

    Figure Lengend Snippet: Crosslinking and precipitation of p75 NTR :Sortilin:proNT3 complexes HEK293 cells transfected with an endocytosis-impaired variant of Sortilin, Sortilin-mut, and p75 NTR as indicated were incubated with or without 50 nM ligand (proNT3 or proNGF) before treatment with crosslinker. Receptor:ligand complexes were immunoprecipitated with anti-p75 NTR , and components were visualised after reducing SDS-PAGE by immunoblotting with anti-Sortilin, anti-proNT3, and anti-p75 NTR . The expression of Sortilin and p75 NTR in input cell lysates were confirmed by immunoblotting. HEK293 human embryonic kidney 293 cells , p75 NTR p75 neurotrophin receptor , Sortilin-mut endocytosis-impaired variant of Sortilin, proNGF proform of nerve growth factor, proNT3 proform of neurotrophin-3, NT3 mature form of neurotrophin-3

    Article Snippet: The blots were blocked in 5% defatted milk in TBST buffer (50mM Tris, 500mM NaCl, 0.1% Tween-20, pH 7.4) for 1 hour at RT and subsequently incubated over night at 4°C in blocking buffer with mouse anti-human Sortilin, rabbit anti-human p75NTR (provided by Moses V. Chao), rabbit anti-proNT3, rabbit anti-NT3 (Alomone Labs Ltd.) or mouse anti-beta-actin (Sigma Aldrich).

    Techniques: Transfection, Variant Assay, Incubation, Immunoprecipitation, SDS Page, Expressing

    Immunoblotting of inner ear tissue for Sortilin, p75 NTR , proNT3 and mature NT3 Subfractions of membranous labyrinth from dissected newborn rats (P3–P5) were pooled, homogenised and centrifuged. The supernatants were subjected to SDS-PAGE followed by immunoblotting for Sortilin, p75 NTR , proNT3 and mature NT3. Beta-actin (~42 kDa) blots are included for evaluation of total protein concentration. (A) Sortilin (~98 kDa) was detected in whole-organ cochleae (Co) and in all in the subfractions i.e. the stria vascularis (I), the organ of Corti with Reissner’s membrane attached (II), the modiolus (not including the spiral ganglion) (III), and the spiral ganglion (IV). (B) P75 NTR (~64 kDa) was detected in the modiolus (III) and the SG (IV). (C) ProNT3 and NT3 were detected in all subfractions. (D) The specificity of proNT3 and NT3 antibodies was tested by immunoblotting of recombinant human proNGF (lane 1 (5 ng) and 2 (1 ng)), proNT3 (lane 3 (5 ng) and 4 (1 ng)), and NT3 (lane 5 (5 ng) and 6 (1 ng)). The proNT3 antibody recognises only proNT3, while the NT3 antibody detects both the proform and the mature form of NT3. kDa kilodalton, Co whole-organ membranous cochlea, p75 NTR p75 neurotrophin receptor, proNT3 proform of neurotrophin-3, NT3 mature form of neurotrophin-3, αSortilin anti-Sortilin, αp75 NTR anti- p75 NTR , αproNT3 anti-proNT3, αNT3 anti-NT3 , αbeta-actin anti-beta actin, P3–P5 postnatal day 3–5

    Journal: The European journal of neuroscience

    Article Title: Proneurotrophin-3 may induce Sortilin dependent death in inner ear neurons

    doi: 10.1111/j.1460-9568.2010.07556.x

    Figure Lengend Snippet: Immunoblotting of inner ear tissue for Sortilin, p75 NTR , proNT3 and mature NT3 Subfractions of membranous labyrinth from dissected newborn rats (P3–P5) were pooled, homogenised and centrifuged. The supernatants were subjected to SDS-PAGE followed by immunoblotting for Sortilin, p75 NTR , proNT3 and mature NT3. Beta-actin (~42 kDa) blots are included for evaluation of total protein concentration. (A) Sortilin (~98 kDa) was detected in whole-organ cochleae (Co) and in all in the subfractions i.e. the stria vascularis (I), the organ of Corti with Reissner’s membrane attached (II), the modiolus (not including the spiral ganglion) (III), and the spiral ganglion (IV). (B) P75 NTR (~64 kDa) was detected in the modiolus (III) and the SG (IV). (C) ProNT3 and NT3 were detected in all subfractions. (D) The specificity of proNT3 and NT3 antibodies was tested by immunoblotting of recombinant human proNGF (lane 1 (5 ng) and 2 (1 ng)), proNT3 (lane 3 (5 ng) and 4 (1 ng)), and NT3 (lane 5 (5 ng) and 6 (1 ng)). The proNT3 antibody recognises only proNT3, while the NT3 antibody detects both the proform and the mature form of NT3. kDa kilodalton, Co whole-organ membranous cochlea, p75 NTR p75 neurotrophin receptor, proNT3 proform of neurotrophin-3, NT3 mature form of neurotrophin-3, αSortilin anti-Sortilin, αp75 NTR anti- p75 NTR , αproNT3 anti-proNT3, αNT3 anti-NT3 , αbeta-actin anti-beta actin, P3–P5 postnatal day 3–5

    Article Snippet: The blots were blocked in 5% defatted milk in TBST buffer (50mM Tris, 500mM NaCl, 0.1% Tween-20, pH 7.4) for 1 hour at RT and subsequently incubated over night at 4°C in blocking buffer with mouse anti-human Sortilin, rabbit anti-human p75NTR (provided by Moses V. Chao), rabbit anti-proNT3, rabbit anti-NT3 (Alomone Labs Ltd.) or mouse anti-beta-actin (Sigma Aldrich).

    Techniques: SDS Page, Protein Concentration, Recombinant

    ProNT3 induced apoptosis in SCG neurons The apoptotic potential of proNT3 was tested in SCG neurons. Here is shown the number of apoptotic neurons in percent of the total number counted. SCG neurons were incubated in growth media containing NGF (20ng/ml), NT3 (2ng/ml), proNGF (2ng/ml) or proNT3 (4ng/ml). After 36 hours, SCG cultures were fixed, immunostained with α-Tuj1, α-Sortilin and DAPI and subjected to apoptosis analysis by morphological evaluation. The results demonstrate that NGF (as opposed to NT3) is imperative for SCG neuron survival. In contrast, both proNGF and proNT3 significantly increased the proportion of apoptotic neurons compared to NGF withdrawal alone. SCG superior cervical ganglion, proNT proneurotrophins, NGF nerve growth factor, proNGF proform of NGF , None no (pro)neurotrophins added , NT3 neurotrophin-3, proNT3 proform of NT3.

    Journal: The European journal of neuroscience

    Article Title: Proneurotrophin-3 may induce Sortilin dependent death in inner ear neurons

    doi: 10.1111/j.1460-9568.2010.07556.x

    Figure Lengend Snippet: ProNT3 induced apoptosis in SCG neurons The apoptotic potential of proNT3 was tested in SCG neurons. Here is shown the number of apoptotic neurons in percent of the total number counted. SCG neurons were incubated in growth media containing NGF (20ng/ml), NT3 (2ng/ml), proNGF (2ng/ml) or proNT3 (4ng/ml). After 36 hours, SCG cultures were fixed, immunostained with α-Tuj1, α-Sortilin and DAPI and subjected to apoptosis analysis by morphological evaluation. The results demonstrate that NGF (as opposed to NT3) is imperative for SCG neuron survival. In contrast, both proNGF and proNT3 significantly increased the proportion of apoptotic neurons compared to NGF withdrawal alone. SCG superior cervical ganglion, proNT proneurotrophins, NGF nerve growth factor, proNGF proform of NGF , None no (pro)neurotrophins added , NT3 neurotrophin-3, proNT3 proform of NT3.

    Article Snippet: The blots were blocked in 5% defatted milk in TBST buffer (50mM Tris, 500mM NaCl, 0.1% Tween-20, pH 7.4) for 1 hour at RT and subsequently incubated over night at 4°C in blocking buffer with mouse anti-human Sortilin, rabbit anti-human p75NTR (provided by Moses V. Chao), rabbit anti-proNT3, rabbit anti-NT3 (Alomone Labs Ltd.) or mouse anti-beta-actin (Sigma Aldrich).

    Techniques: Incubation

    SPR analysis of ligand binding to immobilised Sortilin SPR analysis of receptor-ligand interactions between the NT3 and immobilised Sortilin demonstrated that mature NT3 binds to Sortilin with a Kd ~20 μM (A) . This affinity is significantly lower than the affinity found for the interaction between proNT3 and Sortilin (Kd ~20 nM) (B) , and for the interaction between the receptor and the prodomain sequence of proNT3 fused to GST (Kd~20 nM) (D) . The interaction between Sortilin and proNT3 can be inhibited by the well-known Sortilin ligand neurotensin (C) . NT3 neurotrophin-3, proNT3 proform of NT3, GST-NT3pro prodomain of NT3 coupled to glutathione-s-transferase NTS neurotensin, Kd dissociation constant, SPR Surface plasmon resonance

    Journal: The European journal of neuroscience

    Article Title: Proneurotrophin-3 may induce Sortilin dependent death in inner ear neurons

    doi: 10.1111/j.1460-9568.2010.07556.x

    Figure Lengend Snippet: SPR analysis of ligand binding to immobilised Sortilin SPR analysis of receptor-ligand interactions between the NT3 and immobilised Sortilin demonstrated that mature NT3 binds to Sortilin with a Kd ~20 μM (A) . This affinity is significantly lower than the affinity found for the interaction between proNT3 and Sortilin (Kd ~20 nM) (B) , and for the interaction between the receptor and the prodomain sequence of proNT3 fused to GST (Kd~20 nM) (D) . The interaction between Sortilin and proNT3 can be inhibited by the well-known Sortilin ligand neurotensin (C) . NT3 neurotrophin-3, proNT3 proform of NT3, GST-NT3pro prodomain of NT3 coupled to glutathione-s-transferase NTS neurotensin, Kd dissociation constant, SPR Surface plasmon resonance

    Article Snippet: The blots were blocked in 5% defatted milk in TBST buffer (50mM Tris, 500mM NaCl, 0.1% Tween-20, pH 7.4) for 1 hour at RT and subsequently incubated over night at 4°C in blocking buffer with mouse anti-human Sortilin, rabbit anti-human p75NTR (provided by Moses V. Chao), rabbit anti-proNT3, rabbit anti-NT3 (Alomone Labs Ltd.) or mouse anti-beta-actin (Sigma Aldrich).

    Techniques: SPR Assay, Ligand Binding Assay, Sequencing

    Inhibition of microglial activation after intravitreal injection of recombinant mature NT3 and blockage of P75 NTR . Transgenic mice received twice intravitreal injections of NT3 (0.4 μg), P75 NTR rabbit polyclonal a polyclonal Ab to P75 NTR (1:1 dilution), a combination of both or balanced salt solution ( BSS ) 3d and 6d after tamoxifen (TMX)-induced Muller cell ablation. Eyes were enucleated 10d after TMX treatment for retinal flat-mount staining using an Ab against ionized calcium binding adaptor molecule 1 ( IBA-1 ) for microglia ( A, D, G and J , green ) and peanut-agglutinin ( PNA )-conjugated with Alexa Fluor-594 for cone photoreceptor outer segments ( B, E, H and K , red ). (C, F, I and L) Merged images. (M) Quantitative analysis of IBA-1 stained retinal whole mounts shows intravitreal injection of NT3 and P75 NTR blockage, either alone or in combination, protected photoreceptors concurrently with inhibition of microglial activation. * P

    Journal: Journal of Neuroinflammation

    Article Title: Involvement of NT3 and P75NTR in photoreceptor degeneration following selective M?ller cell ablation

    doi: 10.1186/1742-2094-10-137

    Figure Lengend Snippet: Inhibition of microglial activation after intravitreal injection of recombinant mature NT3 and blockage of P75 NTR . Transgenic mice received twice intravitreal injections of NT3 (0.4 μg), P75 NTR rabbit polyclonal a polyclonal Ab to P75 NTR (1:1 dilution), a combination of both or balanced salt solution ( BSS ) 3d and 6d after tamoxifen (TMX)-induced Muller cell ablation. Eyes were enucleated 10d after TMX treatment for retinal flat-mount staining using an Ab against ionized calcium binding adaptor molecule 1 ( IBA-1 ) for microglia ( A, D, G and J , green ) and peanut-agglutinin ( PNA )-conjugated with Alexa Fluor-594 for cone photoreceptor outer segments ( B, E, H and K , red ). (C, F, I and L) Merged images. (M) Quantitative analysis of IBA-1 stained retinal whole mounts shows intravitreal injection of NT3 and P75 NTR blockage, either alone or in combination, protected photoreceptors concurrently with inhibition of microglial activation. * P

    Article Snippet: Membranes were probed with antibodies to GS (mouse monoclonal, 1:1,000; Millipore, no. MAB302), GFAP (mouse monoclonal, 1:5,000; Neomarker, no. MS-280-P), P75NTR (rabbit polyclonal, a gift from Dr. Moses V. Chao, no. 9651), NT3 (rabbit polyclonal, 1:1,000, Alomone Laboratory, no. ANT-003), pro-NT3 (rabbit polyclonal, 1:500, Alomone Laboratory, no. ANT-012) and rhodopsin (mouse monoclonal, 1:500, Millipore no. MAB5356), guanine nucleotide-binding protein subunit alpha-1 (GNAT1, rabbit polyclonal, 1:500, Santa Cruz no. sc-389) and Gα protein transducin (Gαt, mouse monoclonal, 1:2,000, BD Transduction Laboratories no. 610589).

    Techniques: Inhibition, Activation Assay, Injection, Recombinant, Transgenic Assay, Mouse Assay, Staining, Binding Assay

    Differential expression of mature NT3, pro-NT3, rod and cone phototransduction proteins 7d after Müller cell ablation. (A, F) Western blots using antibodies to detect mature NT3, pro-NT3, rhodopsin, guanine nucleotide-binding protein subunit alpha-1 (GNAT1) and Gα protein transducin (Gαt). GNAT1 and Gαt are proteins essential for rod and cone phototransduction [ 27 - 29 ]. (B-D) Quantitative analysis of protein densitometry showed significant reduction in mature NT3 (B) and upregulation of pro-NT3 (C) , which resulted in a decreased ratio of NT3:pro-NT3 (D) 7d after Müller cell ablation. (E, G, H) The reduced expression of rhodopsin (E) , GNAT1 (G) and Gαt (H) in transgenic (TG) mice indicates that selective Müller cell ablation causes damage to both rod and cone photoreceptors. * P

    Journal: Journal of Neuroinflammation

    Article Title: Involvement of NT3 and P75NTR in photoreceptor degeneration following selective M?ller cell ablation

    doi: 10.1186/1742-2094-10-137

    Figure Lengend Snippet: Differential expression of mature NT3, pro-NT3, rod and cone phototransduction proteins 7d after Müller cell ablation. (A, F) Western blots using antibodies to detect mature NT3, pro-NT3, rhodopsin, guanine nucleotide-binding protein subunit alpha-1 (GNAT1) and Gα protein transducin (Gαt). GNAT1 and Gαt are proteins essential for rod and cone phototransduction [ 27 - 29 ]. (B-D) Quantitative analysis of protein densitometry showed significant reduction in mature NT3 (B) and upregulation of pro-NT3 (C) , which resulted in a decreased ratio of NT3:pro-NT3 (D) 7d after Müller cell ablation. (E, G, H) The reduced expression of rhodopsin (E) , GNAT1 (G) and Gαt (H) in transgenic (TG) mice indicates that selective Müller cell ablation causes damage to both rod and cone photoreceptors. * P

    Article Snippet: Membranes were probed with antibodies to GS (mouse monoclonal, 1:1,000; Millipore, no. MAB302), GFAP (mouse monoclonal, 1:5,000; Neomarker, no. MS-280-P), P75NTR (rabbit polyclonal, a gift from Dr. Moses V. Chao, no. 9651), NT3 (rabbit polyclonal, 1:1,000, Alomone Laboratory, no. ANT-003), pro-NT3 (rabbit polyclonal, 1:500, Alomone Laboratory, no. ANT-012) and rhodopsin (mouse monoclonal, 1:500, Millipore no. MAB5356), guanine nucleotide-binding protein subunit alpha-1 (GNAT1, rabbit polyclonal, 1:500, Santa Cruz no. sc-389) and Gα protein transducin (Gαt, mouse monoclonal, 1:2,000, BD Transduction Laboratories no. 610589).

    Techniques: Expressing, Western Blot, Binding Assay, Transgenic Assay, Mouse Assay

    Intravitreal injection of recombinant NT3 and blockage of P75 NTR protected photoreceptor degeneration. Intravitreal injection was performed in transgenic mice 3d and 6d after tamoxifen induced Müller cell ablation, with one eye receiving the testing reagent and the contralateral eye receiving balanced salt solution (BSS). Doses of injection: (1) NT3, 0.4 μg; (2) P75 NTR rabbit polyclonal Ab (1:1 dilution) and (3) 0.4 μg NT3 + 1:1 dilution of P75 NTR Ab. Eyes were enucleated 10d after TMX treatment for flat-mount staining using fluorescence-conjugated peanut-agglutinin (PNA). (A and B) PNA-labeled cone photoreceptor outer segments ( green ) and nuclear counterstaining in a control ( Ctl ) retina. (C-F) Images from transgenic ( TG ) mice showing changes in cone photoreceptor outer segments and protrusion of photoreceptor nuclei into the subretinal space ( red , indicated by asterisks ) in eyes receiving BSS (C) , NT3 (D) , P75 NTR Ab (E) and a combination of NT3 and P75 NTR Ab (F) . (G) Quantitative analysis of PNA staining showed that intravitreal injection of NT3 and P75 Ab protected the loss of cone photoreceptor outer segments, with a combined treatment more effective than either alone. N = 9 − 11/group. Scale bars in A-F : 100 μm.

    Journal: Journal of Neuroinflammation

    Article Title: Involvement of NT3 and P75NTR in photoreceptor degeneration following selective M?ller cell ablation

    doi: 10.1186/1742-2094-10-137

    Figure Lengend Snippet: Intravitreal injection of recombinant NT3 and blockage of P75 NTR protected photoreceptor degeneration. Intravitreal injection was performed in transgenic mice 3d and 6d after tamoxifen induced Müller cell ablation, with one eye receiving the testing reagent and the contralateral eye receiving balanced salt solution (BSS). Doses of injection: (1) NT3, 0.4 μg; (2) P75 NTR rabbit polyclonal Ab (1:1 dilution) and (3) 0.4 μg NT3 + 1:1 dilution of P75 NTR Ab. Eyes were enucleated 10d after TMX treatment for flat-mount staining using fluorescence-conjugated peanut-agglutinin (PNA). (A and B) PNA-labeled cone photoreceptor outer segments ( green ) and nuclear counterstaining in a control ( Ctl ) retina. (C-F) Images from transgenic ( TG ) mice showing changes in cone photoreceptor outer segments and protrusion of photoreceptor nuclei into the subretinal space ( red , indicated by asterisks ) in eyes receiving BSS (C) , NT3 (D) , P75 NTR Ab (E) and a combination of NT3 and P75 NTR Ab (F) . (G) Quantitative analysis of PNA staining showed that intravitreal injection of NT3 and P75 Ab protected the loss of cone photoreceptor outer segments, with a combined treatment more effective than either alone. N = 9 − 11/group. Scale bars in A-F : 100 μm.

    Article Snippet: Membranes were probed with antibodies to GS (mouse monoclonal, 1:1,000; Millipore, no. MAB302), GFAP (mouse monoclonal, 1:5,000; Neomarker, no. MS-280-P), P75NTR (rabbit polyclonal, a gift from Dr. Moses V. Chao, no. 9651), NT3 (rabbit polyclonal, 1:1,000, Alomone Laboratory, no. ANT-003), pro-NT3 (rabbit polyclonal, 1:500, Alomone Laboratory, no. ANT-012) and rhodopsin (mouse monoclonal, 1:500, Millipore no. MAB5356), guanine nucleotide-binding protein subunit alpha-1 (GNAT1, rabbit polyclonal, 1:500, Santa Cruz no. sc-389) and Gα protein transducin (Gαt, mouse monoclonal, 1:2,000, BD Transduction Laboratories no. 610589).

    Techniques: Injection, Recombinant, Transgenic Assay, Mouse Assay, Staining, Fluorescence, Labeling, CTL Assay

    Inhibition of microglial activation after intravitreal injection of recombinant mature NT3 and blockage of P75 NTR . Transgenic mice received twice intravitreal injections of NT3 (0.4 μg), P75 NTR rabbit polyclonal a polyclonal Ab to P75 NTR (1:1 dilution), a combination of both or balanced salt solution ( BSS ) 3d and 6d after tamoxifen (TMX)-induced Muller cell ablation. Eyes were enucleated 10d after TMX treatment for retinal flat-mount staining using an Ab against ionized calcium binding adaptor molecule 1 ( IBA-1 ) for microglia ( A, D, G and J , green ) and peanut-agglutinin ( PNA )-conjugated with Alexa Fluor-594 for cone photoreceptor outer segments ( B, E, H and K , red ). (C, F, I and L) Merged images. (M) Quantitative analysis of IBA-1 stained retinal whole mounts shows intravitreal injection of NT3 and P75 NTR blockage, either alone or in combination, protected photoreceptors concurrently with inhibition of microglial activation. * P

    Journal: Journal of Neuroinflammation

    Article Title: Involvement of NT3 and P75NTR in photoreceptor degeneration following selective M?ller cell ablation

    doi: 10.1186/1742-2094-10-137

    Figure Lengend Snippet: Inhibition of microglial activation after intravitreal injection of recombinant mature NT3 and blockage of P75 NTR . Transgenic mice received twice intravitreal injections of NT3 (0.4 μg), P75 NTR rabbit polyclonal a polyclonal Ab to P75 NTR (1:1 dilution), a combination of both or balanced salt solution ( BSS ) 3d and 6d after tamoxifen (TMX)-induced Muller cell ablation. Eyes were enucleated 10d after TMX treatment for retinal flat-mount staining using an Ab against ionized calcium binding adaptor molecule 1 ( IBA-1 ) for microglia ( A, D, G and J , green ) and peanut-agglutinin ( PNA )-conjugated with Alexa Fluor-594 for cone photoreceptor outer segments ( B, E, H and K , red ). (C, F, I and L) Merged images. (M) Quantitative analysis of IBA-1 stained retinal whole mounts shows intravitreal injection of NT3 and P75 NTR blockage, either alone or in combination, protected photoreceptors concurrently with inhibition of microglial activation. * P

    Article Snippet: Membranes were probed with antibodies to GS (mouse monoclonal, 1:1,000; Millipore, no. MAB302), GFAP (mouse monoclonal, 1:5,000; Neomarker, no. MS-280-P), P75NTR (rabbit polyclonal, a gift from Dr. Moses V. Chao, no. 9651), NT3 (rabbit polyclonal, 1:1,000, Alomone Laboratory, no. ANT-003), pro-NT3 (rabbit polyclonal, 1:500, Alomone Laboratory, no. ANT-012) and rhodopsin (mouse monoclonal, 1:500, Millipore no. MAB5356), guanine nucleotide-binding protein subunit alpha-1 (GNAT1, rabbit polyclonal, 1:500, Santa Cruz no. sc-389) and Gα protein transducin (Gαt, mouse monoclonal, 1:2,000, BD Transduction Laboratories no. 610589).

    Techniques: Inhibition, Activation Assay, Injection, Recombinant, Transgenic Assay, Mouse Assay, Staining, Binding Assay

    Differential expression of mature NT3, pro-NT3, rod and cone phototransduction proteins 7d after Müller cell ablation. (A, F) Western blots using antibodies to detect mature NT3, pro-NT3, rhodopsin, guanine nucleotide-binding protein subunit alpha-1 (GNAT1) and Gα protein transducin (Gαt). GNAT1 and Gαt are proteins essential for rod and cone phototransduction [ 27 - 29 ]. (B-D) Quantitative analysis of protein densitometry showed significant reduction in mature NT3 (B) and upregulation of pro-NT3 (C) , which resulted in a decreased ratio of NT3:pro-NT3 (D) 7d after Müller cell ablation. (E, G, H) The reduced expression of rhodopsin (E) , GNAT1 (G) and Gαt (H) in transgenic (TG) mice indicates that selective Müller cell ablation causes damage to both rod and cone photoreceptors. * P

    Journal: Journal of Neuroinflammation

    Article Title: Involvement of NT3 and P75NTR in photoreceptor degeneration following selective M?ller cell ablation

    doi: 10.1186/1742-2094-10-137

    Figure Lengend Snippet: Differential expression of mature NT3, pro-NT3, rod and cone phototransduction proteins 7d after Müller cell ablation. (A, F) Western blots using antibodies to detect mature NT3, pro-NT3, rhodopsin, guanine nucleotide-binding protein subunit alpha-1 (GNAT1) and Gα protein transducin (Gαt). GNAT1 and Gαt are proteins essential for rod and cone phototransduction [ 27 - 29 ]. (B-D) Quantitative analysis of protein densitometry showed significant reduction in mature NT3 (B) and upregulation of pro-NT3 (C) , which resulted in a decreased ratio of NT3:pro-NT3 (D) 7d after Müller cell ablation. (E, G, H) The reduced expression of rhodopsin (E) , GNAT1 (G) and Gαt (H) in transgenic (TG) mice indicates that selective Müller cell ablation causes damage to both rod and cone photoreceptors. * P

    Article Snippet: Membranes were probed with antibodies to GS (mouse monoclonal, 1:1,000; Millipore, no. MAB302), GFAP (mouse monoclonal, 1:5,000; Neomarker, no. MS-280-P), P75NTR (rabbit polyclonal, a gift from Dr. Moses V. Chao, no. 9651), NT3 (rabbit polyclonal, 1:1,000, Alomone Laboratory, no. ANT-003), pro-NT3 (rabbit polyclonal, 1:500, Alomone Laboratory, no. ANT-012) and rhodopsin (mouse monoclonal, 1:500, Millipore no. MAB5356), guanine nucleotide-binding protein subunit alpha-1 (GNAT1, rabbit polyclonal, 1:500, Santa Cruz no. sc-389) and Gα protein transducin (Gαt, mouse monoclonal, 1:2,000, BD Transduction Laboratories no. 610589).

    Techniques: Expressing, Western Blot, Binding Assay, Transgenic Assay, Mouse Assay

    Intravitreal injection of recombinant NT3 and blockage of P75 NTR protected photoreceptor degeneration. Intravitreal injection was performed in transgenic mice 3d and 6d after tamoxifen induced Müller cell ablation, with one eye receiving the testing reagent and the contralateral eye receiving balanced salt solution (BSS). Doses of injection: (1) NT3, 0.4 μg; (2) P75 NTR rabbit polyclonal Ab (1:1 dilution) and (3) 0.4 μg NT3 + 1:1 dilution of P75 NTR Ab. Eyes were enucleated 10d after TMX treatment for flat-mount staining using fluorescence-conjugated peanut-agglutinin (PNA). (A and B) PNA-labeled cone photoreceptor outer segments ( green ) and nuclear counterstaining in a control ( Ctl ) retina. (C-F) Images from transgenic ( TG ) mice showing changes in cone photoreceptor outer segments and protrusion of photoreceptor nuclei into the subretinal space ( red , indicated by asterisks ) in eyes receiving BSS (C) , NT3 (D) , P75 NTR Ab (E) and a combination of NT3 and P75 NTR Ab (F) . (G) Quantitative analysis of PNA staining showed that intravitreal injection of NT3 and P75 Ab protected the loss of cone photoreceptor outer segments, with a combined treatment more effective than either alone. N = 9 − 11/group. Scale bars in A-F : 100 μm.

    Journal: Journal of Neuroinflammation

    Article Title: Involvement of NT3 and P75NTR in photoreceptor degeneration following selective M?ller cell ablation

    doi: 10.1186/1742-2094-10-137

    Figure Lengend Snippet: Intravitreal injection of recombinant NT3 and blockage of P75 NTR protected photoreceptor degeneration. Intravitreal injection was performed in transgenic mice 3d and 6d after tamoxifen induced Müller cell ablation, with one eye receiving the testing reagent and the contralateral eye receiving balanced salt solution (BSS). Doses of injection: (1) NT3, 0.4 μg; (2) P75 NTR rabbit polyclonal Ab (1:1 dilution) and (3) 0.4 μg NT3 + 1:1 dilution of P75 NTR Ab. Eyes were enucleated 10d after TMX treatment for flat-mount staining using fluorescence-conjugated peanut-agglutinin (PNA). (A and B) PNA-labeled cone photoreceptor outer segments ( green ) and nuclear counterstaining in a control ( Ctl ) retina. (C-F) Images from transgenic ( TG ) mice showing changes in cone photoreceptor outer segments and protrusion of photoreceptor nuclei into the subretinal space ( red , indicated by asterisks ) in eyes receiving BSS (C) , NT3 (D) , P75 NTR Ab (E) and a combination of NT3 and P75 NTR Ab (F) . (G) Quantitative analysis of PNA staining showed that intravitreal injection of NT3 and P75 Ab protected the loss of cone photoreceptor outer segments, with a combined treatment more effective than either alone. N = 9 − 11/group. Scale bars in A-F : 100 μm.

    Article Snippet: Membranes were probed with antibodies to GS (mouse monoclonal, 1:1,000; Millipore, no. MAB302), GFAP (mouse monoclonal, 1:5,000; Neomarker, no. MS-280-P), P75NTR (rabbit polyclonal, a gift from Dr. Moses V. Chao, no. 9651), NT3 (rabbit polyclonal, 1:1,000, Alomone Laboratory, no. ANT-003), pro-NT3 (rabbit polyclonal, 1:500, Alomone Laboratory, no. ANT-012) and rhodopsin (mouse monoclonal, 1:500, Millipore no. MAB5356), guanine nucleotide-binding protein subunit alpha-1 (GNAT1, rabbit polyclonal, 1:500, Santa Cruz no. sc-389) and Gα protein transducin (Gαt, mouse monoclonal, 1:2,000, BD Transduction Laboratories no. 610589).

    Techniques: Injection, Recombinant, Transgenic Assay, Mouse Assay, Staining, Fluorescence, Labeling, CTL Assay