p75ntr  (Alomone Labs)


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    Alomone Labs p75ntr
    Necdin is coexpressed with TrkA or <t>p75NTR</t> in developing DRGs. A , B , Double immunostaining for necdin and TrkA or p75NTR. Frozen DRG sections of E13.5 mouse embryo were double immunostained for necdin with antibody GN1 (red) and TrkA (or p75NTR) (green), and two images were merged (yellow). Scale bars: A , 50 μm; B , 10 μm. C , Immunoaffinity assay for endogenous necdin-interacting proteins. Tissue lysate (1 mg) of the DRG and spinal cord from E13.5 embryos was applied to the immunoaffinity columns with anti-necdin IgG (NC243) (αNecdin IgG) and control preimmune IgG (Preimmune IgG). The eluates were analyzed by Western blotting for endogenous complexes of necdin with TrkA, p75NTR, and APP. Lysate, Tissue lysate (30 μg).
    P75ntr, 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/p75ntr/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p75ntr - by Bioz Stars, 2022-07
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    Images

    1) Product Images from "Disruption of the Paternal Necdin Gene Diminishes TrkA Signaling for Sensory Neuron Survival"

    Article Title: Disruption of the Paternal Necdin Gene Diminishes TrkA Signaling for Sensory Neuron Survival

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.2083-05.2005

    Necdin is coexpressed with TrkA or p75NTR in developing DRGs. A , B , Double immunostaining for necdin and TrkA or p75NTR. Frozen DRG sections of E13.5 mouse embryo were double immunostained for necdin with antibody GN1 (red) and TrkA (or p75NTR) (green), and two images were merged (yellow). Scale bars: A , 50 μm; B , 10 μm. C , Immunoaffinity assay for endogenous necdin-interacting proteins. Tissue lysate (1 mg) of the DRG and spinal cord from E13.5 embryos was applied to the immunoaffinity columns with anti-necdin IgG (NC243) (αNecdin IgG) and control preimmune IgG (Preimmune IgG). The eluates were analyzed by Western blotting for endogenous complexes of necdin with TrkA, p75NTR, and APP. Lysate, Tissue lysate (30 μg).
    Figure Legend Snippet: Necdin is coexpressed with TrkA or p75NTR in developing DRGs. A , B , Double immunostaining for necdin and TrkA or p75NTR. Frozen DRG sections of E13.5 mouse embryo were double immunostained for necdin with antibody GN1 (red) and TrkA (or p75NTR) (green), and two images were merged (yellow). Scale bars: A , 50 μm; B , 10 μm. C , Immunoaffinity assay for endogenous necdin-interacting proteins. Tissue lysate (1 mg) of the DRG and spinal cord from E13.5 embryos was applied to the immunoaffinity columns with anti-necdin IgG (NC243) (αNecdin IgG) and control preimmune IgG (Preimmune IgG). The eluates were analyzed by Western blotting for endogenous complexes of necdin with TrkA, p75NTR, and APP. Lysate, Tissue lysate (30 μg).

    Techniques Used: Double Immunostaining, Western Blot

    Necdin interacts with both TrkA and p75NTR. A , B , Immunoprecipitation assay for interactions of necdin with TrkA and p75NTR. Lysates of HEK293A cells transfected with expression vectors for FLAG-TrkA, FLAG-p75NTR, FLAG-APP, and necdin were immunoprecipitated (IP) with anti-FLAG antibody M2 (αFLAG) and immunoblotted (IB) with anti-necdin antibody NC243 (αNecdin) ( A , top panel). Conversely, the lysates were immunoprecipitated with antibody NC243 and immunoblotted with antibody M2 ( B , top panel). Expressed proteins in cell lysates are shown in the bottom panels. C , Necdin-enhanced association between TrkA and p75NTR. Lysates of cells transfected with expression vectors for HA-p75NTR (4 μg), FLAG-TrkA (4.5 μg), and necdin (0.1 and 0.5 μg) were immunoprecipitated with antibody M2 and immunoblotted with anti-p75NTR antibody (top panel). D , Interactions of necdin with TrkA mutants. Lysates of cDNA-transfected cells expressing necdin, FLAG-tagged wild-type (WT) TrkA, and the mutants (K547A, Y499A, Y794A) were immunoprecipitated as in A . Phosphorylated Tyr499 in rat TrkA was analyzed using the antibody against phospho-human TrkA (Tyr490) (αpTrkA) ( D , second panel from the top).
    Figure Legend Snippet: Necdin interacts with both TrkA and p75NTR. A , B , Immunoprecipitation assay for interactions of necdin with TrkA and p75NTR. Lysates of HEK293A cells transfected with expression vectors for FLAG-TrkA, FLAG-p75NTR, FLAG-APP, and necdin were immunoprecipitated (IP) with anti-FLAG antibody M2 (αFLAG) and immunoblotted (IB) with anti-necdin antibody NC243 (αNecdin) ( A , top panel). Conversely, the lysates were immunoprecipitated with antibody NC243 and immunoblotted with antibody M2 ( B , top panel). Expressed proteins in cell lysates are shown in the bottom panels. C , Necdin-enhanced association between TrkA and p75NTR. Lysates of cells transfected with expression vectors for HA-p75NTR (4 μg), FLAG-TrkA (4.5 μg), and necdin (0.1 and 0.5 μg) were immunoprecipitated with antibody M2 and immunoblotted with anti-p75NTR antibody (top panel). D , Interactions of necdin with TrkA mutants. Lysates of cDNA-transfected cells expressing necdin, FLAG-tagged wild-type (WT) TrkA, and the mutants (K547A, Y499A, Y794A) were immunoprecipitated as in A . Phosphorylated Tyr499 in rat TrkA was analyzed using the antibody against phospho-human TrkA (Tyr490) (αpTrkA) ( D , second panel from the top).

    Techniques Used: Immunoprecipitation, Transfection, Expressing

    Necdin enhances NGF/TrkA signaling in PC12 cells. A , Coimmunoprecipitation assay for the association between TrkA and p75NTR. PC12 cells infected with recombinant adenoviruses expressing necdin (Ad-Necdin) and β-galactosidase (Ad-LacZ) were treated with NGF (50 ng/ml) for 38 h. Cell lysates (400 μg) were immunoprecipitated (IP) with mouse anti-p75NTR antibody MC192 (αp75NTRm) and immunoblotted (IB) with anti-TrkA antibody (top panel). Expressed proteins in cell lysates are shown in the bottom panels. B , Neurite outgrowth assay. Ad-Necdin- and Ad-LacZ-infected PC12 cells were treated with NGF for 38 h, and cells bearing extended neurites were counted (mean ± SEM; n = 4; p
    Figure Legend Snippet: Necdin enhances NGF/TrkA signaling in PC12 cells. A , Coimmunoprecipitation assay for the association between TrkA and p75NTR. PC12 cells infected with recombinant adenoviruses expressing necdin (Ad-Necdin) and β-galactosidase (Ad-LacZ) were treated with NGF (50 ng/ml) for 38 h. Cell lysates (400 μg) were immunoprecipitated (IP) with mouse anti-p75NTR antibody MC192 (αp75NTRm) and immunoblotted (IB) with anti-TrkA antibody (top panel). Expressed proteins in cell lysates are shown in the bottom panels. B , Neurite outgrowth assay. Ad-Necdin- and Ad-LacZ-infected PC12 cells were treated with NGF for 38 h, and cells bearing extended neurites were counted (mean ± SEM; n = 4; p

    Techniques Used: Co-Immunoprecipitation Assay, Infection, Recombinant, Expressing, Immunoprecipitation, Neurite Outgrowth Assay

    Necdin expression is absent in DRGs of mutant mice lacking the paternal necdin gene. A , The restriction map of mouse Ndn locus. A 7 kb Ndn sequence containing coding (black) and 3′,5′ noncoding (hatched) regions was subcloned into the targeting vector. The 1.5 kb expression unit containing the pgk promoter with Neo r gene (Pgk1/neo r ), polyadenylation signal [Poly(A) signal], was inserted to disrupt the Ndn coding sequence. The vector was introduced to TT2 cells for homologous recombination as described in Materials and Methods. B , Absence of necdin expression in the brain and spinal cord of mice deficient in the paternal necdin gene. Frozen sections of the hypothalamus and spinal cord from E13.5 wild-type (Ndn +m/+p ) and necdin-deficient (Ndn +m/-p ) littermates were immunostained for necdin. HT, Hypothalamus; SC, spinal cord; DBB, diagonal band of Broca; ML, mantle layer. Scale bars: HT, 50 μm; SC, 100 μm. C , Expression patterns of TrkA and p75NTR in necdin-deficient DRGs. Frozen sections of cervical DRGs from E12.5 wild-type (Ndn +m/+p ) and necdin-deficient (Ndn +m/-p ) littermates were immunostained for necdin, TrkA, and p75NTR. Scale bar, 50 μm. D , E , Western blot analysis of necdin, TrkA, p75NTR, and phosphorylated MAPK. Equal amounts of DRG lysates from E12.5 embryos were analyzed by Western blotting. Each lane represents pooled DRGs from a single embryo. Molecular sizes in kilodaltons are shown to the right. E , Signal intensities of TrkA, p75NTR, and pMAPK shown in D were normalized to those of tubulin (TB) and MAPK (mean ± SEM; n = 5 for each protein). * p
    Figure Legend Snippet: Necdin expression is absent in DRGs of mutant mice lacking the paternal necdin gene. A , The restriction map of mouse Ndn locus. A 7 kb Ndn sequence containing coding (black) and 3′,5′ noncoding (hatched) regions was subcloned into the targeting vector. The 1.5 kb expression unit containing the pgk promoter with Neo r gene (Pgk1/neo r ), polyadenylation signal [Poly(A) signal], was inserted to disrupt the Ndn coding sequence. The vector was introduced to TT2 cells for homologous recombination as described in Materials and Methods. B , Absence of necdin expression in the brain and spinal cord of mice deficient in the paternal necdin gene. Frozen sections of the hypothalamus and spinal cord from E13.5 wild-type (Ndn +m/+p ) and necdin-deficient (Ndn +m/-p ) littermates were immunostained for necdin. HT, Hypothalamus; SC, spinal cord; DBB, diagonal band of Broca; ML, mantle layer. Scale bars: HT, 50 μm; SC, 100 μm. C , Expression patterns of TrkA and p75NTR in necdin-deficient DRGs. Frozen sections of cervical DRGs from E12.5 wild-type (Ndn +m/+p ) and necdin-deficient (Ndn +m/-p ) littermates were immunostained for necdin, TrkA, and p75NTR. Scale bar, 50 μm. D , E , Western blot analysis of necdin, TrkA, p75NTR, and phosphorylated MAPK. Equal amounts of DRG lysates from E12.5 embryos were analyzed by Western blotting. Each lane represents pooled DRGs from a single embryo. Molecular sizes in kilodaltons are shown to the right. E , Signal intensities of TrkA, p75NTR, and pMAPK shown in D were normalized to those of tubulin (TB) and MAPK (mean ± SEM; n = 5 for each protein). * p

    Techniques Used: Expressing, Mutagenesis, Mouse Assay, Sequencing, Plasmid Preparation, Homologous Recombination, Western Blot

    2) Product Images from "Differential Hippocampal Expression of BDNF Isoforms and Their Receptors Under Diverse Configurations of the Serotonergic System in a Mice Model of Increased Neuronal Survival"

    Article Title: Differential Hippocampal Expression of BDNF Isoforms and Their Receptors Under Diverse Configurations of the Serotonergic System in a Mice Model of Increased Neuronal Survival

    Journal: Frontiers in Cellular Neuroscience

    doi: 10.3389/fncel.2019.00384

    Effect of 8-OH-DPAT on the protein expression of molecules from the BDNF signaling pathway in the HC, determined by Western Blot. (A) mBDNF. (B) TrkB. (C) proBDNF. (D) p75 in hyposerotonergic (PCPA-treated) or control (vehicle-treated) mice receiving the 5-HT 1 A agonist 8-OH-DPAT (striped bars) or NaCl (white bars). (E) Representative membrane showing signal for p75 (75 kDa) and tubuline (52 kDa). 1: Control-NaCl. 2: Control-DPAT. 3: PCPA-NaCl. 4: PCPA-DPAT. Data are expressed as mean ± S.E.M., n = 6 per experimental group. ∗ p
    Figure Legend Snippet: Effect of 8-OH-DPAT on the protein expression of molecules from the BDNF signaling pathway in the HC, determined by Western Blot. (A) mBDNF. (B) TrkB. (C) proBDNF. (D) p75 in hyposerotonergic (PCPA-treated) or control (vehicle-treated) mice receiving the 5-HT 1 A agonist 8-OH-DPAT (striped bars) or NaCl (white bars). (E) Representative membrane showing signal for p75 (75 kDa) and tubuline (52 kDa). 1: Control-NaCl. 2: Control-DPAT. 3: PCPA-NaCl. 4: PCPA-DPAT. Data are expressed as mean ± S.E.M., n = 6 per experimental group. ∗ p

    Techniques Used: Expressing, Western Blot, Mouse Assay

    Protein expression of molecules from the BDNF signaling pathway determined by Western Blot and representative membranes. (A) mBDNF. (B) TrkB. (C) proBDNF. (D) p75 in hyposerotonergic (PCPA-treated) or control (vehicle-treated) mice for 4 weeks. (E–H) Representative membrane showing signal for TrkB, p75, pro-BDNF, and mBDNF in Control and PCPA-treatd mice. Data are expressed as mean ± S.E.M., n = 11 (control) and 12 (PCPA). ∗ p
    Figure Legend Snippet: Protein expression of molecules from the BDNF signaling pathway determined by Western Blot and representative membranes. (A) mBDNF. (B) TrkB. (C) proBDNF. (D) p75 in hyposerotonergic (PCPA-treated) or control (vehicle-treated) mice for 4 weeks. (E–H) Representative membrane showing signal for TrkB, p75, pro-BDNF, and mBDNF in Control and PCPA-treatd mice. Data are expressed as mean ± S.E.M., n = 11 (control) and 12 (PCPA). ∗ p

    Techniques Used: Expressing, Western Blot, Mouse Assay

    3) Product Images from "Influence of bacterial components on the developmental programming of enteric neurons, et al. Influence of bacterial components on the developmental programming of enteric neurons"

    Article Title: Influence of bacterial components on the developmental programming of enteric neurons, et al. Influence of bacterial components on the developmental programming of enteric neurons

    Journal: Physiological Reports

    doi: 10.14814/phy2.14611

    Characterization of ENCDC cultures. Flow cytometry of ENCDC cultures stained with antibodies against the immature neuronal marker, p75 NTR+ ‐FITC demonstrated 95.1% p75 NTR+ ‐positive, 97.3% viability in subculture 3, and 96.1% p75 NTR+ ‐positive, 98.2% viability in subculture 4 (a). Proportion of ENCDC cultures characterized for expression of p75NTR+, pH3, 5‐HT, nNOS, and TH. Scale bar = 20 µm (b). There was a significant increase in the proportion of serotonergic neurons and nitrergic neurons in subculture 5 (gray) compared to subculture 4 (black), but no significant difference in proliferating cells or dopaminergic neurons across the cultures. *p ≤ .05. Values are presented as mean ± SEM (c)
    Figure Legend Snippet: Characterization of ENCDC cultures. Flow cytometry of ENCDC cultures stained with antibodies against the immature neuronal marker, p75 NTR+ ‐FITC demonstrated 95.1% p75 NTR+ ‐positive, 97.3% viability in subculture 3, and 96.1% p75 NTR+ ‐positive, 98.2% viability in subculture 4 (a). Proportion of ENCDC cultures characterized for expression of p75NTR+, pH3, 5‐HT, nNOS, and TH. Scale bar = 20 µm (b). There was a significant increase in the proportion of serotonergic neurons and nitrergic neurons in subculture 5 (gray) compared to subculture 4 (black), but no significant difference in proliferating cells or dopaminergic neurons across the cultures. *p ≤ .05. Values are presented as mean ± SEM (c)

    Techniques Used: Flow Cytometry, Staining, Marker, Expressing

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    Alomone Labs p75ntr
    Necdin is coexpressed with TrkA or <t>p75NTR</t> in developing DRGs. A , B , Double immunostaining for necdin and TrkA or p75NTR. Frozen DRG sections of E13.5 mouse embryo were double immunostained for necdin with antibody GN1 (red) and TrkA (or p75NTR) (green), and two images were merged (yellow). Scale bars: A , 50 μm; B , 10 μm. C , Immunoaffinity assay for endogenous necdin-interacting proteins. Tissue lysate (1 mg) of the DRG and spinal cord from E13.5 embryos was applied to the immunoaffinity columns with anti-necdin IgG (NC243) (αNecdin IgG) and control preimmune IgG (Preimmune IgG). The eluates were analyzed by Western blotting for endogenous complexes of necdin with TrkA, p75NTR, and APP. Lysate, Tissue lysate (30 μg).
    P75ntr, 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/p75ntr/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p75ntr - by Bioz Stars, 2022-07
    93/100 stars
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    Necdin is coexpressed with TrkA or p75NTR in developing DRGs. A , B , Double immunostaining for necdin and TrkA or p75NTR. Frozen DRG sections of E13.5 mouse embryo were double immunostained for necdin with antibody GN1 (red) and TrkA (or p75NTR) (green), and two images were merged (yellow). Scale bars: A , 50 μm; B , 10 μm. C , Immunoaffinity assay for endogenous necdin-interacting proteins. Tissue lysate (1 mg) of the DRG and spinal cord from E13.5 embryos was applied to the immunoaffinity columns with anti-necdin IgG (NC243) (αNecdin IgG) and control preimmune IgG (Preimmune IgG). The eluates were analyzed by Western blotting for endogenous complexes of necdin with TrkA, p75NTR, and APP. Lysate, Tissue lysate (30 μg).

    Journal: The Journal of Neuroscience

    Article Title: Disruption of the Paternal Necdin Gene Diminishes TrkA Signaling for Sensory Neuron Survival

    doi: 10.1523/JNEUROSCI.2083-05.2005

    Figure Lengend Snippet: Necdin is coexpressed with TrkA or p75NTR in developing DRGs. A , B , Double immunostaining for necdin and TrkA or p75NTR. Frozen DRG sections of E13.5 mouse embryo were double immunostained for necdin with antibody GN1 (red) and TrkA (or p75NTR) (green), and two images were merged (yellow). Scale bars: A , 50 μm; B , 10 μm. C , Immunoaffinity assay for endogenous necdin-interacting proteins. Tissue lysate (1 mg) of the DRG and spinal cord from E13.5 embryos was applied to the immunoaffinity columns with anti-necdin IgG (NC243) (αNecdin IgG) and control preimmune IgG (Preimmune IgG). The eluates were analyzed by Western blotting for endogenous complexes of necdin with TrkA, p75NTR, and APP. Lysate, Tissue lysate (30 μg).

    Article Snippet: Antibodies used for coimmunoprecipitation assay are as follows: FLAG (M2; 1:50; Sigma), Necdin (NC243; 1:50), and p75NTR (MC192; 1:10; Alomone Labs, Jerusalem, Israel).

    Techniques: Double Immunostaining, Western Blot

    Necdin interacts with both TrkA and p75NTR. A , B , Immunoprecipitation assay for interactions of necdin with TrkA and p75NTR. Lysates of HEK293A cells transfected with expression vectors for FLAG-TrkA, FLAG-p75NTR, FLAG-APP, and necdin were immunoprecipitated (IP) with anti-FLAG antibody M2 (αFLAG) and immunoblotted (IB) with anti-necdin antibody NC243 (αNecdin) ( A , top panel). Conversely, the lysates were immunoprecipitated with antibody NC243 and immunoblotted with antibody M2 ( B , top panel). Expressed proteins in cell lysates are shown in the bottom panels. C , Necdin-enhanced association between TrkA and p75NTR. Lysates of cells transfected with expression vectors for HA-p75NTR (4 μg), FLAG-TrkA (4.5 μg), and necdin (0.1 and 0.5 μg) were immunoprecipitated with antibody M2 and immunoblotted with anti-p75NTR antibody (top panel). D , Interactions of necdin with TrkA mutants. Lysates of cDNA-transfected cells expressing necdin, FLAG-tagged wild-type (WT) TrkA, and the mutants (K547A, Y499A, Y794A) were immunoprecipitated as in A . Phosphorylated Tyr499 in rat TrkA was analyzed using the antibody against phospho-human TrkA (Tyr490) (αpTrkA) ( D , second panel from the top).

    Journal: The Journal of Neuroscience

    Article Title: Disruption of the Paternal Necdin Gene Diminishes TrkA Signaling for Sensory Neuron Survival

    doi: 10.1523/JNEUROSCI.2083-05.2005

    Figure Lengend Snippet: Necdin interacts with both TrkA and p75NTR. A , B , Immunoprecipitation assay for interactions of necdin with TrkA and p75NTR. Lysates of HEK293A cells transfected with expression vectors for FLAG-TrkA, FLAG-p75NTR, FLAG-APP, and necdin were immunoprecipitated (IP) with anti-FLAG antibody M2 (αFLAG) and immunoblotted (IB) with anti-necdin antibody NC243 (αNecdin) ( A , top panel). Conversely, the lysates were immunoprecipitated with antibody NC243 and immunoblotted with antibody M2 ( B , top panel). Expressed proteins in cell lysates are shown in the bottom panels. C , Necdin-enhanced association between TrkA and p75NTR. Lysates of cells transfected with expression vectors for HA-p75NTR (4 μg), FLAG-TrkA (4.5 μg), and necdin (0.1 and 0.5 μg) were immunoprecipitated with antibody M2 and immunoblotted with anti-p75NTR antibody (top panel). D , Interactions of necdin with TrkA mutants. Lysates of cDNA-transfected cells expressing necdin, FLAG-tagged wild-type (WT) TrkA, and the mutants (K547A, Y499A, Y794A) were immunoprecipitated as in A . Phosphorylated Tyr499 in rat TrkA was analyzed using the antibody against phospho-human TrkA (Tyr490) (αpTrkA) ( D , second panel from the top).

    Article Snippet: Antibodies used for coimmunoprecipitation assay are as follows: FLAG (M2; 1:50; Sigma), Necdin (NC243; 1:50), and p75NTR (MC192; 1:10; Alomone Labs, Jerusalem, Israel).

    Techniques: Immunoprecipitation, Transfection, Expressing

    Necdin enhances NGF/TrkA signaling in PC12 cells. A , Coimmunoprecipitation assay for the association between TrkA and p75NTR. PC12 cells infected with recombinant adenoviruses expressing necdin (Ad-Necdin) and β-galactosidase (Ad-LacZ) were treated with NGF (50 ng/ml) for 38 h. Cell lysates (400 μg) were immunoprecipitated (IP) with mouse anti-p75NTR antibody MC192 (αp75NTRm) and immunoblotted (IB) with anti-TrkA antibody (top panel). Expressed proteins in cell lysates are shown in the bottom panels. B , Neurite outgrowth assay. Ad-Necdin- and Ad-LacZ-infected PC12 cells were treated with NGF for 38 h, and cells bearing extended neurites were counted (mean ± SEM; n = 4; p

    Journal: The Journal of Neuroscience

    Article Title: Disruption of the Paternal Necdin Gene Diminishes TrkA Signaling for Sensory Neuron Survival

    doi: 10.1523/JNEUROSCI.2083-05.2005

    Figure Lengend Snippet: Necdin enhances NGF/TrkA signaling in PC12 cells. A , Coimmunoprecipitation assay for the association between TrkA and p75NTR. PC12 cells infected with recombinant adenoviruses expressing necdin (Ad-Necdin) and β-galactosidase (Ad-LacZ) were treated with NGF (50 ng/ml) for 38 h. Cell lysates (400 μg) were immunoprecipitated (IP) with mouse anti-p75NTR antibody MC192 (αp75NTRm) and immunoblotted (IB) with anti-TrkA antibody (top panel). Expressed proteins in cell lysates are shown in the bottom panels. B , Neurite outgrowth assay. Ad-Necdin- and Ad-LacZ-infected PC12 cells were treated with NGF for 38 h, and cells bearing extended neurites were counted (mean ± SEM; n = 4; p

    Article Snippet: Antibodies used for coimmunoprecipitation assay are as follows: FLAG (M2; 1:50; Sigma), Necdin (NC243; 1:50), and p75NTR (MC192; 1:10; Alomone Labs, Jerusalem, Israel).

    Techniques: Co-Immunoprecipitation Assay, Infection, Recombinant, Expressing, Immunoprecipitation, Neurite Outgrowth Assay

    Necdin expression is absent in DRGs of mutant mice lacking the paternal necdin gene. A , The restriction map of mouse Ndn locus. A 7 kb Ndn sequence containing coding (black) and 3′,5′ noncoding (hatched) regions was subcloned into the targeting vector. The 1.5 kb expression unit containing the pgk promoter with Neo r gene (Pgk1/neo r ), polyadenylation signal [Poly(A) signal], was inserted to disrupt the Ndn coding sequence. The vector was introduced to TT2 cells for homologous recombination as described in Materials and Methods. B , Absence of necdin expression in the brain and spinal cord of mice deficient in the paternal necdin gene. Frozen sections of the hypothalamus and spinal cord from E13.5 wild-type (Ndn +m/+p ) and necdin-deficient (Ndn +m/-p ) littermates were immunostained for necdin. HT, Hypothalamus; SC, spinal cord; DBB, diagonal band of Broca; ML, mantle layer. Scale bars: HT, 50 μm; SC, 100 μm. C , Expression patterns of TrkA and p75NTR in necdin-deficient DRGs. Frozen sections of cervical DRGs from E12.5 wild-type (Ndn +m/+p ) and necdin-deficient (Ndn +m/-p ) littermates were immunostained for necdin, TrkA, and p75NTR. Scale bar, 50 μm. D , E , Western blot analysis of necdin, TrkA, p75NTR, and phosphorylated MAPK. Equal amounts of DRG lysates from E12.5 embryos were analyzed by Western blotting. Each lane represents pooled DRGs from a single embryo. Molecular sizes in kilodaltons are shown to the right. E , Signal intensities of TrkA, p75NTR, and pMAPK shown in D were normalized to those of tubulin (TB) and MAPK (mean ± SEM; n = 5 for each protein). * p

    Journal: The Journal of Neuroscience

    Article Title: Disruption of the Paternal Necdin Gene Diminishes TrkA Signaling for Sensory Neuron Survival

    doi: 10.1523/JNEUROSCI.2083-05.2005

    Figure Lengend Snippet: Necdin expression is absent in DRGs of mutant mice lacking the paternal necdin gene. A , The restriction map of mouse Ndn locus. A 7 kb Ndn sequence containing coding (black) and 3′,5′ noncoding (hatched) regions was subcloned into the targeting vector. The 1.5 kb expression unit containing the pgk promoter with Neo r gene (Pgk1/neo r ), polyadenylation signal [Poly(A) signal], was inserted to disrupt the Ndn coding sequence. The vector was introduced to TT2 cells for homologous recombination as described in Materials and Methods. B , Absence of necdin expression in the brain and spinal cord of mice deficient in the paternal necdin gene. Frozen sections of the hypothalamus and spinal cord from E13.5 wild-type (Ndn +m/+p ) and necdin-deficient (Ndn +m/-p ) littermates were immunostained for necdin. HT, Hypothalamus; SC, spinal cord; DBB, diagonal band of Broca; ML, mantle layer. Scale bars: HT, 50 μm; SC, 100 μm. C , Expression patterns of TrkA and p75NTR in necdin-deficient DRGs. Frozen sections of cervical DRGs from E12.5 wild-type (Ndn +m/+p ) and necdin-deficient (Ndn +m/-p ) littermates were immunostained for necdin, TrkA, and p75NTR. Scale bar, 50 μm. D , E , Western blot analysis of necdin, TrkA, p75NTR, and phosphorylated MAPK. Equal amounts of DRG lysates from E12.5 embryos were analyzed by Western blotting. Each lane represents pooled DRGs from a single embryo. Molecular sizes in kilodaltons are shown to the right. E , Signal intensities of TrkA, p75NTR, and pMAPK shown in D were normalized to those of tubulin (TB) and MAPK (mean ± SEM; n = 5 for each protein). * p

    Article Snippet: Antibodies used for coimmunoprecipitation assay are as follows: FLAG (M2; 1:50; Sigma), Necdin (NC243; 1:50), and p75NTR (MC192; 1:10; Alomone Labs, Jerusalem, Israel).

    Techniques: Expressing, Mutagenesis, Mouse Assay, Sequencing, Plasmid Preparation, Homologous Recombination, Western Blot