anti-rabbit Search Results


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  • 99
    Vector Laboratories anti rabbit antibody
    Anti Rabbit Antibody, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 99/100, based on 482 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti rabbit antibody/product/Vector Laboratories
    Average 99 stars, based on 482 article reviews
    Price from $9.99 to $1999.99
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    99
    Thermo Fisher anti rabbit antibody
    Cell surface nucleolin is expressed in angiogenic blood vessels. Balb/c nu/nu mice were subcutaneously injected with matrigel supplemented with bFGF. 8 d later, an antinucleolin antibody (NCL3) or control IgG was injected into the tail vein of the mice. The matrigel plugs were removed 1 h after the injection, sectioned, and examined for the presence of rabbit IgG using Alexa-594 <t>anti–rabbit</t> IgG (red). Blood vessels were stained with anti-CD31 antibody (green), and nuclei were counterstained with DAPI (blue). The injected NCL3 colocalizes the blood vessel staining in the matrigel plugs (a), but no injected rabbit IgG is detected in the plugs (e). No specific NCL3 accumulation over the IgG control is seen in any of the tissues examined: b and f, skin; c and g, heart; or d and h, brain. b–d, NCL3; f–h, IgG.
    Anti Rabbit Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1495 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti rabbit antibody/product/Thermo Fisher
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    91
    Cell Signaling Technology Inc rabbit anti
    Dopamine D1R stimulation activates <t>Fyn</t> but not <t>Src</t> in the DMS but not in the DLS or NAc. (A) Schematic representation of DMS, DLS, and NAc dissection. (B–D) SKF81297 (SKF, 5 mg/kg) or vehicle (2% DMSO) was systemically administered (i.p.). The DMS (B) , DLS (C) , and NAc (D) were collected 15 min after drug administration. The level of Tyr420 phosphorylation (pFyn) (upper panel), Fyn (middle panel), and GAPDH, which was used as a loading control (bottom panel) were determined by western blot analysis. Data are presented as the densitometry value of the pFyn divided by the densitometry values of Fyn ± SEM and expressed as % of vehicle. (E,F) Mouse striatal sections were treated with SKF81297 (SKF, 10 μM) or vehicle (0.1% DMSO) for 15 min. The DMS was dissected and Fyn or Src were immunoprecipitated using anti-Fyn (IP Fyn) (E) or anti-Src (IP Src) (F) antibodies. Mouse IgG was used as a negative control (IP IgG). Fyn and Src immunoreactivity (lower panels) as well as Tyr417/420[Src/Fyn] phosphorylation (upper panels) were determined by westernblot analysis. Protein levels in the total lysates were measured in parallel (Input). Two-tailed t -test. (B–D) *** p
    Rabbit Anti, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 91/100, based on 233 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti/product/Cell Signaling Technology Inc
    Average 91 stars, based on 233 article reviews
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    90
    Biotium anti rabbit ab
    Dopamine D1R stimulation activates <t>Fyn</t> but not <t>Src</t> in the DMS but not in the DLS or NAc. (A) Schematic representation of DMS, DLS, and NAc dissection. (B–D) SKF81297 (SKF, 5 mg/kg) or vehicle (2% DMSO) was systemically administered (i.p.). The DMS (B) , DLS (C) , and NAc (D) were collected 15 min after drug administration. The level of Tyr420 phosphorylation (pFyn) (upper panel), Fyn (middle panel), and GAPDH, which was used as a loading control (bottom panel) were determined by western blot analysis. Data are presented as the densitometry value of the pFyn divided by the densitometry values of Fyn ± SEM and expressed as % of vehicle. (E,F) Mouse striatal sections were treated with SKF81297 (SKF, 10 μM) or vehicle (0.1% DMSO) for 15 min. The DMS was dissected and Fyn or Src were immunoprecipitated using anti-Fyn (IP Fyn) (E) or anti-Src (IP Src) (F) antibodies. Mouse IgG was used as a negative control (IP IgG). Fyn and Src immunoreactivity (lower panels) as well as Tyr417/420[Src/Fyn] phosphorylation (upper panels) were determined by westernblot analysis. Protein levels in the total lysates were measured in parallel (Input). Two-tailed t -test. (B–D) *** p
    Anti Rabbit Ab, supplied by Biotium, used in various techniques. Bioz Stars score: 90/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti rabbit ab/product/Biotium
    Average 90 stars, based on 4 article reviews
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    93
    Santa Cruz Biotechnology anti rabbit antibodies
    Dopamine D1R stimulation activates <t>Fyn</t> but not <t>Src</t> in the DMS but not in the DLS or NAc. (A) Schematic representation of DMS, DLS, and NAc dissection. (B–D) SKF81297 (SKF, 5 mg/kg) or vehicle (2% DMSO) was systemically administered (i.p.). The DMS (B) , DLS (C) , and NAc (D) were collected 15 min after drug administration. The level of Tyr420 phosphorylation (pFyn) (upper panel), Fyn (middle panel), and GAPDH, which was used as a loading control (bottom panel) were determined by western blot analysis. Data are presented as the densitometry value of the pFyn divided by the densitometry values of Fyn ± SEM and expressed as % of vehicle. (E,F) Mouse striatal sections were treated with SKF81297 (SKF, 10 μM) or vehicle (0.1% DMSO) for 15 min. The DMS was dissected and Fyn or Src were immunoprecipitated using anti-Fyn (IP Fyn) (E) or anti-Src (IP Src) (F) antibodies. Mouse IgG was used as a negative control (IP IgG). Fyn and Src immunoreactivity (lower panels) as well as Tyr417/420[Src/Fyn] phosphorylation (upper panels) were determined by westernblot analysis. Protein levels in the total lysates were measured in parallel (Input). Two-tailed t -test. (B–D) *** p
    Anti Rabbit Antibodies, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 817 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti rabbit antibodies/product/Santa Cruz Biotechnology
    Average 93 stars, based on 817 article reviews
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    85
    Jackson Immuno anti rabbit cy3 ab
    Dopamine D1R stimulation activates <t>Fyn</t> but not <t>Src</t> in the DMS but not in the DLS or NAc. (A) Schematic representation of DMS, DLS, and NAc dissection. (B–D) SKF81297 (SKF, 5 mg/kg) or vehicle (2% DMSO) was systemically administered (i.p.). The DMS (B) , DLS (C) , and NAc (D) were collected 15 min after drug administration. The level of Tyr420 phosphorylation (pFyn) (upper panel), Fyn (middle panel), and GAPDH, which was used as a loading control (bottom panel) were determined by western blot analysis. Data are presented as the densitometry value of the pFyn divided by the densitometry values of Fyn ± SEM and expressed as % of vehicle. (E,F) Mouse striatal sections were treated with SKF81297 (SKF, 10 μM) or vehicle (0.1% DMSO) for 15 min. The DMS was dissected and Fyn or Src were immunoprecipitated using anti-Fyn (IP Fyn) (E) or anti-Src (IP Src) (F) antibodies. Mouse IgG was used as a negative control (IP IgG). Fyn and Src immunoreactivity (lower panels) as well as Tyr417/420[Src/Fyn] phosphorylation (upper panels) were determined by westernblot analysis. Protein levels in the total lysates were measured in parallel (Input). Two-tailed t -test. (B–D) *** p
    Anti Rabbit Cy3 Ab, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 85/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti rabbit cy3 ab/product/Jackson Immuno
    Average 85 stars, based on 7 article reviews
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    90
    Thermo Fisher anti rabbit pe abs
    Dopamine D1R stimulation activates <t>Fyn</t> but not <t>Src</t> in the DMS but not in the DLS or NAc. (A) Schematic representation of DMS, DLS, and NAc dissection. (B–D) SKF81297 (SKF, 5 mg/kg) or vehicle (2% DMSO) was systemically administered (i.p.). The DMS (B) , DLS (C) , and NAc (D) were collected 15 min after drug administration. The level of Tyr420 phosphorylation (pFyn) (upper panel), Fyn (middle panel), and GAPDH, which was used as a loading control (bottom panel) were determined by western blot analysis. Data are presented as the densitometry value of the pFyn divided by the densitometry values of Fyn ± SEM and expressed as % of vehicle. (E,F) Mouse striatal sections were treated with SKF81297 (SKF, 10 μM) or vehicle (0.1% DMSO) for 15 min. The DMS was dissected and Fyn or Src were immunoprecipitated using anti-Fyn (IP Fyn) (E) or anti-Src (IP Src) (F) antibodies. Mouse IgG was used as a negative control (IP IgG). Fyn and Src immunoreactivity (lower panels) as well as Tyr417/420[Src/Fyn] phosphorylation (upper panels) were determined by westernblot analysis. Protein levels in the total lysates were measured in parallel (Input). Two-tailed t -test. (B–D) *** p
    Anti Rabbit Pe Abs, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Santa Cruz Biotechnology rabbit polyclonal ab
    Antitumor effect of P 4 N-induced antiserum in immunodeficient mice and the target antigens of the antiserum. ( A ) Nude mice bearing CT26 cell tumors ( n = 5 per group) were treated with 100 μL of PBS, PBS antisera, or P 4 N antisera weekly. Tumor volumes were measured every 2 d after treatment. ( B ) Titers of specific anti-CT26 or JC cell antibodies for the PBS antisera or P 4 N antisera on days 0, 17, 24, and 31 were measured. ( C ) Effects of P 4 N on the changes in the isotypes of specific anti-CT26 cell antibodies were examined. The titers of these antisera (1,600× dilution) were measured for IgM, IgG1, IgG2a, IgG2b, or IgA. ( D ) IHC analysis of antisera bound to tumor antigens on the surface of CT26. Color development by <t>HRP-conjugated</t> anti-mouse antibodies indicates the antisera antibody. ( E ) Antisera binding to tumor antigens on the surface of CT26 cells was indirectly detected with FITC-conjugated goat anti-mouse <t>IgG</t> antibody. ( F ) Subcellular location of antigens recognized by the antisera was monitored by confocal microscopy. Alexa Fluor 568-conjugated anti-mouse Ig antibodies were used to display the presence of antisera (red). Alexa Fluor 488-Con A and DAPI were used to indicate the cell membrane (green) and the cell nucleus (blue), respectively. ( G ) Normal sera, PBS antisera, or P 4 N anti-sera were used to probe Western blots of membrane proteins of extracted from CT26 cells. Two bands of 78 kDa and 55 kDa were visualized: E-cadherin proteins were used as a loading control. For all results, significant differences between the P 4 N antisera group and the PBS antisera group were calculated and labeled with * P
    Rabbit Polyclonal Ab, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 340 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Cell Signaling Technology Inc anti rabbit polyclonal ab
    Antitumor effect of P 4 N-induced antiserum in immunodeficient mice and the target antigens of the antiserum. ( A ) Nude mice bearing CT26 cell tumors ( n = 5 per group) were treated with 100 μL of PBS, PBS antisera, or P 4 N antisera weekly. Tumor volumes were measured every 2 d after treatment. ( B ) Titers of specific anti-CT26 or JC cell antibodies for the PBS antisera or P 4 N antisera on days 0, 17, 24, and 31 were measured. ( C ) Effects of P 4 N on the changes in the isotypes of specific anti-CT26 cell antibodies were examined. The titers of these antisera (1,600× dilution) were measured for IgM, IgG1, IgG2a, IgG2b, or IgA. ( D ) IHC analysis of antisera bound to tumor antigens on the surface of CT26. Color development by <t>HRP-conjugated</t> anti-mouse antibodies indicates the antisera antibody. ( E ) Antisera binding to tumor antigens on the surface of CT26 cells was indirectly detected with FITC-conjugated goat anti-mouse <t>IgG</t> antibody. ( F ) Subcellular location of antigens recognized by the antisera was monitored by confocal microscopy. Alexa Fluor 568-conjugated anti-mouse Ig antibodies were used to display the presence of antisera (red). Alexa Fluor 488-Con A and DAPI were used to indicate the cell membrane (green) and the cell nucleus (blue), respectively. ( G ) Normal sera, PBS antisera, or P 4 N anti-sera were used to probe Western blots of membrane proteins of extracted from CT26 cells. Two bands of 78 kDa and 55 kDa were visualized: E-cadherin proteins were used as a loading control. For all results, significant differences between the P 4 N antisera group and the PBS antisera group were calculated and labeled with * P
    Anti Rabbit Polyclonal Ab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 85/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 85 stars, based on 15 article reviews
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    90
    Enzo Biochem anti rabbit secondary ab
    Antitumor effect of P 4 N-induced antiserum in immunodeficient mice and the target antigens of the antiserum. ( A ) Nude mice bearing CT26 cell tumors ( n = 5 per group) were treated with 100 μL of PBS, PBS antisera, or P 4 N antisera weekly. Tumor volumes were measured every 2 d after treatment. ( B ) Titers of specific anti-CT26 or JC cell antibodies for the PBS antisera or P 4 N antisera on days 0, 17, 24, and 31 were measured. ( C ) Effects of P 4 N on the changes in the isotypes of specific anti-CT26 cell antibodies were examined. The titers of these antisera (1,600× dilution) were measured for IgM, IgG1, IgG2a, IgG2b, or IgA. ( D ) IHC analysis of antisera bound to tumor antigens on the surface of CT26. Color development by <t>HRP-conjugated</t> anti-mouse antibodies indicates the antisera antibody. ( E ) Antisera binding to tumor antigens on the surface of CT26 cells was indirectly detected with FITC-conjugated goat anti-mouse <t>IgG</t> antibody. ( F ) Subcellular location of antigens recognized by the antisera was monitored by confocal microscopy. Alexa Fluor 568-conjugated anti-mouse Ig antibodies were used to display the presence of antisera (red). Alexa Fluor 488-Con A and DAPI were used to indicate the cell membrane (green) and the cell nucleus (blue), respectively. ( G ) Normal sera, PBS antisera, or P 4 N anti-sera were used to probe Western blots of membrane proteins of extracted from CT26 cells. Two bands of 78 kDa and 55 kDa were visualized: E-cadherin proteins were used as a loading control. For all results, significant differences between the P 4 N antisera group and the PBS antisera group were calculated and labeled with * P
    Anti Rabbit Secondary Ab, supplied by Enzo Biochem, used in various techniques. Bioz Stars score: 90/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    GE Healthcare anti rabbit ig antibody
    Reactivity of polyclonal antibody against actinin-4 peptide. ( A ) Immunoblot analysis reveals that the polyclonal antibody raised against actinin-4 peptide (lane  1 ) reacts with a single  protein of ∼100 kD in the cell lysate of WiDr cells. A blot with  normal rabbit IgG (lane  2 ) is included as a negative control. Molecular masses (in kD) are shown on the left. ( B ) In vitro translation products of actinin-4 (lanes  1  and  3 ) and actinin-1 (lanes  2   and  4 ) were immunoprecipitated by the anti–actinin-4 polyclonal  antibody (lanes  1  and  2 ) or normal rabbit IgG (lanes  3  and  4 ).  SDS-PAGE and autoradiography reveal that this polyclonal antibody is reactive with actinin-4 protein, but not with actinin-1. ( C )  Confocal immunofluorescence microscopy showing the subcellular localization of actinin-4 in lung fibroblasts MRC-5. Arrow indicates the nuclear staining, and arrowheads indicate linear staining of actinin-4 along actin stress fibers. Bar, 5 μm.
    Anti Rabbit Ig Antibody, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 88/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Jackson Immuno anti rabbit percp ab
    Reactivity of polyclonal antibody against actinin-4 peptide. ( A ) Immunoblot analysis reveals that the polyclonal antibody raised against actinin-4 peptide (lane  1 ) reacts with a single  protein of ∼100 kD in the cell lysate of WiDr cells. A blot with  normal rabbit IgG (lane  2 ) is included as a negative control. Molecular masses (in kD) are shown on the left. ( B ) In vitro translation products of actinin-4 (lanes  1  and  3 ) and actinin-1 (lanes  2   and  4 ) were immunoprecipitated by the anti–actinin-4 polyclonal  antibody (lanes  1  and  2 ) or normal rabbit IgG (lanes  3  and  4 ).  SDS-PAGE and autoradiography reveal that this polyclonal antibody is reactive with actinin-4 protein, but not with actinin-1. ( C )  Confocal immunofluorescence microscopy showing the subcellular localization of actinin-4 in lung fibroblasts MRC-5. Arrow indicates the nuclear staining, and arrowheads indicate linear staining of actinin-4 along actin stress fibers. Bar, 5 μm.
    Anti Rabbit Percp Ab, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Santa Cruz Biotechnology anti rabbit s6k1 ab
    GLI1 protein levels are unchanged following <t>S6K1</t> knockdown. Western blot analysis of GLI1 and S6K1 protein expression in SK-N-AS cells, following siRNA-mediated knockdown of GLI1 and S6K1. Note the reduction of the GLI1 and S6K1 protein bands by the siRNAs targeting GLI1 (siGLI1) and S6K1 (siS6), respectively. siCN indicates the control siRNA treatment and β-Actin was used as the endogenous protein control. Quantitation of protein expression, using the ImageJ software, is shown in the bar graphs. Each bar represents the mean ± SEM of triplicate values from a representative experiment. *, Statistical significant, P
    Anti Rabbit S6k1 Ab, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 86/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Santa Cruz Biotechnology anti rabbit hrp antibodies
    GLI1 protein levels are unchanged following <t>S6K1</t> knockdown. Western blot analysis of GLI1 and S6K1 protein expression in SK-N-AS cells, following siRNA-mediated knockdown of GLI1 and S6K1. Note the reduction of the GLI1 and S6K1 protein bands by the siRNAs targeting GLI1 (siGLI1) and S6K1 (siS6), respectively. siCN indicates the control siRNA treatment and β-Actin was used as the endogenous protein control. Quantitation of protein expression, using the ImageJ software, is shown in the bar graphs. Each bar represents the mean ± SEM of triplicate values from a representative experiment. *, Statistical significant, P
    Anti Rabbit Hrp Antibodies, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 89/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    GE Healthcare anti rabbit immunoglobulin g antibodies
    The death domain of IRAK1 interacts with CaMKII. (A) LCL lysates were immunoprecipitated with either normal <t>IgG</t> or anti-IRAK1 antibodies and analyzed by CaMKIIγ and IRAK1 Western blotting. (B) Schematic representation of IRAK1 WT and IRAK1 DD
    Anti Rabbit Immunoglobulin G Antibodies, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 88/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 88 stars, based on 23 article reviews
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    80
    Agilent technologies swine anti rabbit ab
    The death domain of IRAK1 interacts with CaMKII. (A) LCL lysates were immunoprecipitated with either normal <t>IgG</t> or anti-IRAK1 antibodies and analyzed by CaMKIIγ and IRAK1 Western blotting. (B) Schematic representation of IRAK1 WT and IRAK1 DD
    Swine Anti Rabbit Ab, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 80/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Rockland Immunochemicals anti rabbit cy2 secondary ab
    The death domain of IRAK1 interacts with CaMKII. (A) LCL lysates were immunoprecipitated with either normal <t>IgG</t> or anti-IRAK1 antibodies and analyzed by CaMKIIγ and IRAK1 Western blotting. (B) Schematic representation of IRAK1 WT and IRAK1 DD
    Anti Rabbit Cy2 Secondary Ab, supplied by Rockland Immunochemicals, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Covance anti rabbit p75ntr ab
    Induction of <t>p75NTR</t> and proNGF in the white matter (corpus callosum) after CCI-TBI. ( A–D ) Immunoblot analysis showed that both p75NTR and proNGF protein levels are increased after TBI in the ipsilateral white matter as compared to uninjured (UN) and sham injured tissue. Two-way ANOVA, ( A ) for p75NTR: side, F (1,12) = 92.42, P
    Anti Rabbit P75ntr Ab, supplied by Covance, used in various techniques. Bioz Stars score: 90/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher secondary anti rabbit ab af488
    Induction of <t>p75NTR</t> and proNGF in the white matter (corpus callosum) after CCI-TBI. ( A–D ) Immunoblot analysis showed that both p75NTR and proNGF protein levels are increased after TBI in the ipsilateral white matter as compared to uninjured (UN) and sham injured tissue. Two-way ANOVA, ( A ) for p75NTR: side, F (1,12) = 92.42, P
    Secondary Anti Rabbit Ab Af488, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Abcam anti rabbit secondary antibody
    Induction of <t>p75NTR</t> and proNGF in the white matter (corpus callosum) after CCI-TBI. ( A–D ) Immunoblot analysis showed that both p75NTR and proNGF protein levels are increased after TBI in the ipsilateral white matter as compared to uninjured (UN) and sham injured tissue. Two-way ANOVA, ( A ) for p75NTR: side, F (1,12) = 92.42, P
    Anti Rabbit Secondary Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 548 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Abcam rabbit polyclonal αsma ab
    Induction of <t>p75NTR</t> and proNGF in the white matter (corpus callosum) after CCI-TBI. ( A–D ) Immunoblot analysis showed that both p75NTR and proNGF protein levels are increased after TBI in the ipsilateral white matter as compared to uninjured (UN) and sham injured tissue. Two-way ANOVA, ( A ) for p75NTR: side, F (1,12) = 92.42, P
    Rabbit Polyclonal αsma Ab, supplied by Abcam, used in various techniques. Bioz Stars score: 88/100, based on 42 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Thermo Fisher anti rabbit alexa 594 antibody
    Induction of <t>p75NTR</t> and proNGF in the white matter (corpus callosum) after CCI-TBI. ( A–D ) Immunoblot analysis showed that both p75NTR and proNGF protein levels are increased after TBI in the ipsilateral white matter as compared to uninjured (UN) and sham injured tissue. Two-way ANOVA, ( A ) for p75NTR: side, F (1,12) = 92.42, P
    Anti Rabbit Alexa 594 Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell surface nucleolin is expressed in angiogenic blood vessels. Balb/c nu/nu mice were subcutaneously injected with matrigel supplemented with bFGF. 8 d later, an antinucleolin antibody (NCL3) or control IgG was injected into the tail vein of the mice. The matrigel plugs were removed 1 h after the injection, sectioned, and examined for the presence of rabbit IgG using Alexa-594 anti–rabbit IgG (red). Blood vessels were stained with anti-CD31 antibody (green), and nuclei were counterstained with DAPI (blue). The injected NCL3 colocalizes the blood vessel staining in the matrigel plugs (a), but no injected rabbit IgG is detected in the plugs (e). No specific NCL3 accumulation over the IgG control is seen in any of the tissues examined: b and f, skin; c and g, heart; or d and h, brain. b–d, NCL3; f–h, IgG.

    Journal: The Journal of Cell Biology

    Article Title: Nucleolin expressed at the cell surface is a marker of endothelial cells in angiogenic blood vessels

    doi: 10.1083/jcb.200304132

    Figure Lengend Snippet: Cell surface nucleolin is expressed in angiogenic blood vessels. Balb/c nu/nu mice were subcutaneously injected with matrigel supplemented with bFGF. 8 d later, an antinucleolin antibody (NCL3) or control IgG was injected into the tail vein of the mice. The matrigel plugs were removed 1 h after the injection, sectioned, and examined for the presence of rabbit IgG using Alexa-594 anti–rabbit IgG (red). Blood vessels were stained with anti-CD31 antibody (green), and nuclei were counterstained with DAPI (blue). The injected NCL3 colocalizes the blood vessel staining in the matrigel plugs (a), but no injected rabbit IgG is detected in the plugs (e). No specific NCL3 accumulation over the IgG control is seen in any of the tissues examined: b and f, skin; c and g, heart; or d and h, brain. b–d, NCL3; f–h, IgG.

    Article Snippet: Bound antibodies were detected with Alexa-594–labeled anti–rabbit antibody (Molecular Probes) and visualized by fluorescence microscopy.

    Techniques: Mouse Assay, Injection, Staining

    Antinucleolin antibodies inhibit F3 internalization by cells. Exponentially growing MDA-MB-435 cells were incubated with 1 μM FITC-F3 or FITC-Tat peptide for 2 h at 37°C. FITC-F3 is internalized and transported into the nucleus (a, FITC-F3, green; b, red channel; c, merge). Coincubation with antinucleolin antibody NCL3 inhibits the cellular uptake and subsequent nuclear transport of the peptide (e, F3-FITC, green; f, NCL3, red; g, merge). NCL2 has no influence on uptake of F3 (i, F3-FITC, green; j, NCL2, red; k, merge). Internalization of FITC-Tat peptide (d) is not affected by NCL3 (h) or NCL2 (l). The antibodies were detected with Alexa-594 anti–rabbit IgG (red). Nuclei were stained with DAPI (blue). The images were obtained by confocal microscopy. Bars, 10 μm.

    Journal: The Journal of Cell Biology

    Article Title: Nucleolin expressed at the cell surface is a marker of endothelial cells in angiogenic blood vessels

    doi: 10.1083/jcb.200304132

    Figure Lengend Snippet: Antinucleolin antibodies inhibit F3 internalization by cells. Exponentially growing MDA-MB-435 cells were incubated with 1 μM FITC-F3 or FITC-Tat peptide for 2 h at 37°C. FITC-F3 is internalized and transported into the nucleus (a, FITC-F3, green; b, red channel; c, merge). Coincubation with antinucleolin antibody NCL3 inhibits the cellular uptake and subsequent nuclear transport of the peptide (e, F3-FITC, green; f, NCL3, red; g, merge). NCL2 has no influence on uptake of F3 (i, F3-FITC, green; j, NCL2, red; k, merge). Internalization of FITC-Tat peptide (d) is not affected by NCL3 (h) or NCL2 (l). The antibodies were detected with Alexa-594 anti–rabbit IgG (red). Nuclei were stained with DAPI (blue). The images were obtained by confocal microscopy. Bars, 10 μm.

    Article Snippet: Bound antibodies were detected with Alexa-594–labeled anti–rabbit antibody (Molecular Probes) and visualized by fluorescence microscopy.

    Techniques: Multiple Displacement Amplification, Incubation, Staining, Confocal Microscopy

    Intravenously injected antinucleolin antibody accumulates in tumor blood vessels. An affinity-purified rabbit antinucleolin antibody (NCL3) was injected into the tail vein of mice bearing MDA-MB-435 xenograft tumors. The tumor and various organs were removed 1 h after the injection, sectioned, and examined for the presence of rabbit IgG using Alexa-594 anti–rabbit IgG (red). Blood vessels were stained with anti-CD31 antibody (green), and nuclei were counterstained with DAPI (blue). The antinucleolin antibody has bound to tumor blood vessels (a and b), but is not seen in the skin (c). Rabbit IgG injected similarly as a control does not bind to tumor blood vessels (d).

    Journal: The Journal of Cell Biology

    Article Title: Nucleolin expressed at the cell surface is a marker of endothelial cells in angiogenic blood vessels

    doi: 10.1083/jcb.200304132

    Figure Lengend Snippet: Intravenously injected antinucleolin antibody accumulates in tumor blood vessels. An affinity-purified rabbit antinucleolin antibody (NCL3) was injected into the tail vein of mice bearing MDA-MB-435 xenograft tumors. The tumor and various organs were removed 1 h after the injection, sectioned, and examined for the presence of rabbit IgG using Alexa-594 anti–rabbit IgG (red). Blood vessels were stained with anti-CD31 antibody (green), and nuclei were counterstained with DAPI (blue). The antinucleolin antibody has bound to tumor blood vessels (a and b), but is not seen in the skin (c). Rabbit IgG injected similarly as a control does not bind to tumor blood vessels (d).

    Article Snippet: Bound antibodies were detected with Alexa-594–labeled anti–rabbit antibody (Molecular Probes) and visualized by fluorescence microscopy.

    Techniques: Injection, Affinity Purification, Mouse Assay, Multiple Displacement Amplification, Staining

    Dopamine D1R stimulation activates Fyn but not Src in the DMS but not in the DLS or NAc. (A) Schematic representation of DMS, DLS, and NAc dissection. (B–D) SKF81297 (SKF, 5 mg/kg) or vehicle (2% DMSO) was systemically administered (i.p.). The DMS (B) , DLS (C) , and NAc (D) were collected 15 min after drug administration. The level of Tyr420 phosphorylation (pFyn) (upper panel), Fyn (middle panel), and GAPDH, which was used as a loading control (bottom panel) were determined by western blot analysis. Data are presented as the densitometry value of the pFyn divided by the densitometry values of Fyn ± SEM and expressed as % of vehicle. (E,F) Mouse striatal sections were treated with SKF81297 (SKF, 10 μM) or vehicle (0.1% DMSO) for 15 min. The DMS was dissected and Fyn or Src were immunoprecipitated using anti-Fyn (IP Fyn) (E) or anti-Src (IP Src) (F) antibodies. Mouse IgG was used as a negative control (IP IgG). Fyn and Src immunoreactivity (lower panels) as well as Tyr417/420[Src/Fyn] phosphorylation (upper panels) were determined by westernblot analysis. Protein levels in the total lysates were measured in parallel (Input). Two-tailed t -test. (B–D) *** p

    Journal: Frontiers in Molecular Neuroscience

    Article Title: Fyn Signaling Is Compartmentalized to Dopamine D1 Receptor Expressing Neurons in the Dorsal Medial Striatum

    doi: 10.3389/fnmol.2017.00273

    Figure Lengend Snippet: Dopamine D1R stimulation activates Fyn but not Src in the DMS but not in the DLS or NAc. (A) Schematic representation of DMS, DLS, and NAc dissection. (B–D) SKF81297 (SKF, 5 mg/kg) or vehicle (2% DMSO) was systemically administered (i.p.). The DMS (B) , DLS (C) , and NAc (D) were collected 15 min after drug administration. The level of Tyr420 phosphorylation (pFyn) (upper panel), Fyn (middle panel), and GAPDH, which was used as a loading control (bottom panel) were determined by western blot analysis. Data are presented as the densitometry value of the pFyn divided by the densitometry values of Fyn ± SEM and expressed as % of vehicle. (E,F) Mouse striatal sections were treated with SKF81297 (SKF, 10 μM) or vehicle (0.1% DMSO) for 15 min. The DMS was dissected and Fyn or Src were immunoprecipitated using anti-Fyn (IP Fyn) (E) or anti-Src (IP Src) (F) antibodies. Mouse IgG was used as a negative control (IP IgG). Fyn and Src immunoreactivity (lower panels) as well as Tyr417/420[Src/Fyn] phosphorylation (upper panels) were determined by westernblot analysis. Protein levels in the total lysates were measured in parallel (Input). Two-tailed t -test. (B–D) *** p

    Article Snippet: Rabbit anti-[pY1472]GluN2B antibodies and rabbit anti-[pY418/420]Src/Fyn antibodies were purchased from Cell Signaling Technology (Beverly, MA).

    Techniques: Dissection, Western Blot, Immunoprecipitation, Negative Control, Two Tailed Test

    Antitumor effect of P 4 N-induced antiserum in immunodeficient mice and the target antigens of the antiserum. ( A ) Nude mice bearing CT26 cell tumors ( n = 5 per group) were treated with 100 μL of PBS, PBS antisera, or P 4 N antisera weekly. Tumor volumes were measured every 2 d after treatment. ( B ) Titers of specific anti-CT26 or JC cell antibodies for the PBS antisera or P 4 N antisera on days 0, 17, 24, and 31 were measured. ( C ) Effects of P 4 N on the changes in the isotypes of specific anti-CT26 cell antibodies were examined. The titers of these antisera (1,600× dilution) were measured for IgM, IgG1, IgG2a, IgG2b, or IgA. ( D ) IHC analysis of antisera bound to tumor antigens on the surface of CT26. Color development by HRP-conjugated anti-mouse antibodies indicates the antisera antibody. ( E ) Antisera binding to tumor antigens on the surface of CT26 cells was indirectly detected with FITC-conjugated goat anti-mouse IgG antibody. ( F ) Subcellular location of antigens recognized by the antisera was monitored by confocal microscopy. Alexa Fluor 568-conjugated anti-mouse Ig antibodies were used to display the presence of antisera (red). Alexa Fluor 488-Con A and DAPI were used to indicate the cell membrane (green) and the cell nucleus (blue), respectively. ( G ) Normal sera, PBS antisera, or P 4 N anti-sera were used to probe Western blots of membrane proteins of extracted from CT26 cells. Two bands of 78 kDa and 55 kDa were visualized: E-cadherin proteins were used as a loading control. For all results, significant differences between the P 4 N antisera group and the PBS antisera group were calculated and labeled with * P

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: In vivo amelioration of endogenous antitumor autoantibodies via low-dose P4N through the LTA4H/activin A/BAFF pathway

    doi: 10.1073/pnas.1604752113

    Figure Lengend Snippet: Antitumor effect of P 4 N-induced antiserum in immunodeficient mice and the target antigens of the antiserum. ( A ) Nude mice bearing CT26 cell tumors ( n = 5 per group) were treated with 100 μL of PBS, PBS antisera, or P 4 N antisera weekly. Tumor volumes were measured every 2 d after treatment. ( B ) Titers of specific anti-CT26 or JC cell antibodies for the PBS antisera or P 4 N antisera on days 0, 17, 24, and 31 were measured. ( C ) Effects of P 4 N on the changes in the isotypes of specific anti-CT26 cell antibodies were examined. The titers of these antisera (1,600× dilution) were measured for IgM, IgG1, IgG2a, IgG2b, or IgA. ( D ) IHC analysis of antisera bound to tumor antigens on the surface of CT26. Color development by HRP-conjugated anti-mouse antibodies indicates the antisera antibody. ( E ) Antisera binding to tumor antigens on the surface of CT26 cells was indirectly detected with FITC-conjugated goat anti-mouse IgG antibody. ( F ) Subcellular location of antigens recognized by the antisera was monitored by confocal microscopy. Alexa Fluor 568-conjugated anti-mouse Ig antibodies were used to display the presence of antisera (red). Alexa Fluor 488-Con A and DAPI were used to indicate the cell membrane (green) and the cell nucleus (blue), respectively. ( G ) Normal sera, PBS antisera, or P 4 N anti-sera were used to probe Western blots of membrane proteins of extracted from CT26 cells. Two bands of 78 kDa and 55 kDa were visualized: E-cadherin proteins were used as a loading control. For all results, significant differences between the P 4 N antisera group and the PBS antisera group were calculated and labeled with * P

    Article Snippet: The sections were probed with rat monoclonal anti-F4/80 (1:50 dilution; GeneTex) and anti-CD20 (1:200 dilution; Santa Cruz Biotechnology), hamster monoclonal anti-CD11c (1:25 dilution; GeneTex), rabbit monoclonal anti-CD3 (1:50 dilution; GeneTex), and rabbit polyclonal anti–TNF-α and anti–IL-8 antibodies (1:50 dilution; Assay BioTech) at 4 °C overnight, and the detection antibodies were recognized using a horseradish peroxidase (HRP)-conjugated anti-rat, anti-hamster, or anti-rabbit IgG antibody (1:1,500 dilution; Santa Cruz Biotechnology).

    Techniques: Mouse Assay, Immunohistochemistry, Binding Assay, Confocal Microscopy, Western Blot, Labeling

    Reactivity of polyclonal antibody against actinin-4 peptide. ( A ) Immunoblot analysis reveals that the polyclonal antibody raised against actinin-4 peptide (lane  1 ) reacts with a single  protein of ∼100 kD in the cell lysate of WiDr cells. A blot with  normal rabbit IgG (lane  2 ) is included as a negative control. Molecular masses (in kD) are shown on the left. ( B ) In vitro translation products of actinin-4 (lanes  1  and  3 ) and actinin-1 (lanes  2   and  4 ) were immunoprecipitated by the anti–actinin-4 polyclonal  antibody (lanes  1  and  2 ) or normal rabbit IgG (lanes  3  and  4 ).  SDS-PAGE and autoradiography reveal that this polyclonal antibody is reactive with actinin-4 protein, but not with actinin-1. ( C )  Confocal immunofluorescence microscopy showing the subcellular localization of actinin-4 in lung fibroblasts MRC-5. Arrow indicates the nuclear staining, and arrowheads indicate linear staining of actinin-4 along actin stress fibers. Bar, 5 μm.

    Journal: The Journal of Cell Biology

    Article Title: Actinin-4, a Novel Actin-bundling Protein Associated with Cell Motility and Cancer Invasion

    doi:

    Figure Lengend Snippet: Reactivity of polyclonal antibody against actinin-4 peptide. ( A ) Immunoblot analysis reveals that the polyclonal antibody raised against actinin-4 peptide (lane 1 ) reacts with a single protein of ∼100 kD in the cell lysate of WiDr cells. A blot with normal rabbit IgG (lane 2 ) is included as a negative control. Molecular masses (in kD) are shown on the left. ( B ) In vitro translation products of actinin-4 (lanes 1 and 3 ) and actinin-1 (lanes 2 and 4 ) were immunoprecipitated by the anti–actinin-4 polyclonal antibody (lanes 1 and 2 ) or normal rabbit IgG (lanes 3 and 4 ). SDS-PAGE and autoradiography reveal that this polyclonal antibody is reactive with actinin-4 protein, but not with actinin-1. ( C ) Confocal immunofluorescence microscopy showing the subcellular localization of actinin-4 in lung fibroblasts MRC-5. Arrow indicates the nuclear staining, and arrowheads indicate linear staining of actinin-4 along actin stress fibers. Bar, 5 μm.

    Article Snippet: After incubation with antibodies at 4°C overnight, the blots were detected using a horseradish peroxidase–conjugated anti–mouse IgG+IgM (IBL, Fujioka, Japan) or anti–rabbit Ig antibody and ECL Western blotting detection reagents ( Amersham International ), as instructed by the suppliers.

    Techniques: Negative Control, In Vitro, Immunoprecipitation, SDS Page, Autoradiography, Immunofluorescence, Microscopy, Staining

    GLI1 protein levels are unchanged following S6K1 knockdown. Western blot analysis of GLI1 and S6K1 protein expression in SK-N-AS cells, following siRNA-mediated knockdown of GLI1 and S6K1. Note the reduction of the GLI1 and S6K1 protein bands by the siRNAs targeting GLI1 (siGLI1) and S6K1 (siS6), respectively. siCN indicates the control siRNA treatment and β-Actin was used as the endogenous protein control. Quantitation of protein expression, using the ImageJ software, is shown in the bar graphs. Each bar represents the mean ± SEM of triplicate values from a representative experiment. *, Statistical significant, P

    Journal: BMC Cancer

    Article Title: The impact of S6K1 kinase on neuroblastoma cell proliferation is independent of GLI1 signaling

    doi: 10.1186/1471-2407-14-600

    Figure Lengend Snippet: GLI1 protein levels are unchanged following S6K1 knockdown. Western blot analysis of GLI1 and S6K1 protein expression in SK-N-AS cells, following siRNA-mediated knockdown of GLI1 and S6K1. Note the reduction of the GLI1 and S6K1 protein bands by the siRNAs targeting GLI1 (siGLI1) and S6K1 (siS6), respectively. siCN indicates the control siRNA treatment and β-Actin was used as the endogenous protein control. Quantitation of protein expression, using the ImageJ software, is shown in the bar graphs. Each bar represents the mean ± SEM of triplicate values from a representative experiment. *, Statistical significant, P

    Article Snippet: The membrane was incubated at 4°C overnight in 5% skim milk in TBST (Tris Buffered Saline with Tween 20) with anti-rabbit GLI1 Ab (#2553, Cell Signaling) or anti-rabbit S6K1 Ab (sc-230, Santa Cruz Biotechnology) followed by incubation with goat anti-rabbit secondary antibodies for 1 hour in 5% skim milk in TBST and visualized using chemiluminescent substrate (Thermo Scientific).

    Techniques: Western Blot, Expressing, Quantitation Assay, Software

    SK-N-AS cellular proliferation is insensitive to S6K1 or GLI1 overexpression. (A) SK-N-AS cells, cultured for 48 hours following transfection with control pCMV5 vector, and expression constructs for wild type S6K1 (S6K1 WT) constitutively activated S6K1 (S6K1T389E), function-loss S6K1 (S6K1T389A) and GLI1, were subjected to the EdU incorporation assay for 4 hours. (B) SK-N-AS cells, cultured for 48 hours following transfection with control siRNAs and pCMV5 vector (siCN + pCMV), S6K1 siRNAs and pCMV5 vector (siS6K1 + pCMV) and S6K1 siRNAs and GLI1 expression construct (siS6K1 + pGLI1), were subjected to the EdU incorporation assay for 4 hours. The data were analyzed with the one-way ANOVA test followed by Tukey’s multiple comparison using the GraphPad Prism software. Each bar represents the mean ± SEM of three independent experiments *, Statistical significant, P

    Journal: BMC Cancer

    Article Title: The impact of S6K1 kinase on neuroblastoma cell proliferation is independent of GLI1 signaling

    doi: 10.1186/1471-2407-14-600

    Figure Lengend Snippet: SK-N-AS cellular proliferation is insensitive to S6K1 or GLI1 overexpression. (A) SK-N-AS cells, cultured for 48 hours following transfection with control pCMV5 vector, and expression constructs for wild type S6K1 (S6K1 WT) constitutively activated S6K1 (S6K1T389E), function-loss S6K1 (S6K1T389A) and GLI1, were subjected to the EdU incorporation assay for 4 hours. (B) SK-N-AS cells, cultured for 48 hours following transfection with control siRNAs and pCMV5 vector (siCN + pCMV), S6K1 siRNAs and pCMV5 vector (siS6K1 + pCMV) and S6K1 siRNAs and GLI1 expression construct (siS6K1 + pGLI1), were subjected to the EdU incorporation assay for 4 hours. The data were analyzed with the one-way ANOVA test followed by Tukey’s multiple comparison using the GraphPad Prism software. Each bar represents the mean ± SEM of three independent experiments *, Statistical significant, P

    Article Snippet: The membrane was incubated at 4°C overnight in 5% skim milk in TBST (Tris Buffered Saline with Tween 20) with anti-rabbit GLI1 Ab (#2553, Cell Signaling) or anti-rabbit S6K1 Ab (sc-230, Santa Cruz Biotechnology) followed by incubation with goat anti-rabbit secondary antibodies for 1 hour in 5% skim milk in TBST and visualized using chemiluminescent substrate (Thermo Scientific).

    Techniques: Over Expression, Cell Culture, Transfection, Plasmid Preparation, Expressing, Construct, Software

    S6K1 and GLI1 knockdown reduces SK-N-AS cellular proliferation. SK-N-AS cells, cultured for 48 hours following transfection with control (siCN), GLI1 (siGLI1) or S6K1 (siS6K1) siRNAs, were subjected to the EdU incorporation assay for 4 hours. The percentage of cells labeled with Alexa Fluor 488 azide was detected by flow cytometry. The data were analyzed with the one-way ANOVA test followed by Tukey’s multiple comparison using the GraphPad Prism software. Each bar represents the mean ± SEM of three independent experiments. *, Statistical significant, P

    Journal: BMC Cancer

    Article Title: The impact of S6K1 kinase on neuroblastoma cell proliferation is independent of GLI1 signaling

    doi: 10.1186/1471-2407-14-600

    Figure Lengend Snippet: S6K1 and GLI1 knockdown reduces SK-N-AS cellular proliferation. SK-N-AS cells, cultured for 48 hours following transfection with control (siCN), GLI1 (siGLI1) or S6K1 (siS6K1) siRNAs, were subjected to the EdU incorporation assay for 4 hours. The percentage of cells labeled with Alexa Fluor 488 azide was detected by flow cytometry. The data were analyzed with the one-way ANOVA test followed by Tukey’s multiple comparison using the GraphPad Prism software. Each bar represents the mean ± SEM of three independent experiments. *, Statistical significant, P

    Article Snippet: The membrane was incubated at 4°C overnight in 5% skim milk in TBST (Tris Buffered Saline with Tween 20) with anti-rabbit GLI1 Ab (#2553, Cell Signaling) or anti-rabbit S6K1 Ab (sc-230, Santa Cruz Biotechnology) followed by incubation with goat anti-rabbit secondary antibodies for 1 hour in 5% skim milk in TBST and visualized using chemiluminescent substrate (Thermo Scientific).

    Techniques: Cell Culture, Transfection, Labeling, Flow Cytometry, Cytometry, Software

    GLI1 but not S6K1 knockdown reduces GLI1, GLI2, GLI3, SMO and PTCH2 expression. The expression of S6K1 (A) , GLI1 (B) , GLI2 (C) , GLI3 (D) , the signaling molecule SMO (E) and the typical GLI1 target gene PTCH2 (F) in SK-N-AS cells, following siRNA knockdown of GLI1 and S6K1, was determined by real-time PCR. Data are represented as relative expression (2 -∆∆Ct values), calculated by subtracting the Ct value of the housekeeping gene TBP from the Ct value of the interrogated transcripts (∆Ct), and normalized to the ∆Ct value obtained with siCN. Error bars indicate the standard deviation. *, Statistical significant, P

    Journal: BMC Cancer

    Article Title: The impact of S6K1 kinase on neuroblastoma cell proliferation is independent of GLI1 signaling

    doi: 10.1186/1471-2407-14-600

    Figure Lengend Snippet: GLI1 but not S6K1 knockdown reduces GLI1, GLI2, GLI3, SMO and PTCH2 expression. The expression of S6K1 (A) , GLI1 (B) , GLI2 (C) , GLI3 (D) , the signaling molecule SMO (E) and the typical GLI1 target gene PTCH2 (F) in SK-N-AS cells, following siRNA knockdown of GLI1 and S6K1, was determined by real-time PCR. Data are represented as relative expression (2 -∆∆Ct values), calculated by subtracting the Ct value of the housekeeping gene TBP from the Ct value of the interrogated transcripts (∆Ct), and normalized to the ∆Ct value obtained with siCN. Error bars indicate the standard deviation. *, Statistical significant, P

    Article Snippet: The membrane was incubated at 4°C overnight in 5% skim milk in TBST (Tris Buffered Saline with Tween 20) with anti-rabbit GLI1 Ab (#2553, Cell Signaling) or anti-rabbit S6K1 Ab (sc-230, Santa Cruz Biotechnology) followed by incubation with goat anti-rabbit secondary antibodies for 1 hour in 5% skim milk in TBST and visualized using chemiluminescent substrate (Thermo Scientific).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Standard Deviation

    The death domain of IRAK1 interacts with CaMKII. (A) LCL lysates were immunoprecipitated with either normal IgG or anti-IRAK1 antibodies and analyzed by CaMKIIγ and IRAK1 Western blotting. (B) Schematic representation of IRAK1 WT and IRAK1 DD

    Journal: Molecular and Cellular Biology

    Article Title: Role of Ca2+/Calmodulin-Dependent Kinase II–IRAK1 Interaction in LMP1-Induced NF-κB Activation

    doi: 10.1128/MCB.00912-13

    Figure Lengend Snippet: The death domain of IRAK1 interacts with CaMKII. (A) LCL lysates were immunoprecipitated with either normal IgG or anti-IRAK1 antibodies and analyzed by CaMKIIγ and IRAK1 Western blotting. (B) Schematic representation of IRAK1 WT and IRAK1 DD

    Article Snippet: Enhanced chemiluminescence detection reagents (Pierce, Rockford, IL) and secondary peroxidase-labeled anti-mouse or anti-rabbit immunoglobulin G antibodies (Amersham Biosciences, Piscataway, NJ) were used according to the manufacturer's directions.

    Techniques: Immunoprecipitation, Western Blot

    Induction of p75NTR and proNGF in the white matter (corpus callosum) after CCI-TBI. ( A–D ) Immunoblot analysis showed that both p75NTR and proNGF protein levels are increased after TBI in the ipsilateral white matter as compared to uninjured (UN) and sham injured tissue. Two-way ANOVA, ( A ) for p75NTR: side, F (1,12) = 92.42, P

    Journal: Brain

    Article Title: A novel inhibitor of p75-neurotrophin receptor improves functional outcomes in two models of traumatic brain injury

    doi: 10.1093/brain/aww074

    Figure Lengend Snippet: Induction of p75NTR and proNGF in the white matter (corpus callosum) after CCI-TBI. ( A–D ) Immunoblot analysis showed that both p75NTR and proNGF protein levels are increased after TBI in the ipsilateral white matter as compared to uninjured (UN) and sham injured tissue. Two-way ANOVA, ( A ) for p75NTR: side, F (1,12) = 92.42, P

    Article Snippet: Anti-rabbit p75NTR Ab (1:1000 dilution; COVANCE) was added and incubated overnight at 4 °C.

    Techniques:

    Acute expression of p75NTR after FPI-TBI and reduction of neurodegeneration with EVT901 in vivo . ( A ) Immunohistochemical expression of p75NTR at 1–2 days in the dentate gyrus of TBI rats using specific anti-p75NTR antibody. ( B ) Representative photomicrographs (original magnification ×10) of Fluoro-Jade® staining of damaged neurons in the granular layer of the dentate gyrus in FPI-TBI rats 1 week post-injury. ( C ) Representative photomicrographs of Fluoro-Jade®-stained sections from the thalamus 14-days post-trauma (ipsilateral versus contralateral to the injury). ( D ) Quantification of Fluoro-Jade® B-positive degenerating cells in the thalamus of FPI-TBI rats after EVT901 treatment. Following Fluoro-Jade staining, the digital images were collected and the damaged cells were quantified using Explora Nova Software. Three mg/kg/day of EVT901 protected neurons in the thalamus after TBI [one-way ANOVA, EVT901 effect, F (3,18) = 8.46, P = 0.001]. # P

    Journal: Brain

    Article Title: A novel inhibitor of p75-neurotrophin receptor improves functional outcomes in two models of traumatic brain injury

    doi: 10.1093/brain/aww074

    Figure Lengend Snippet: Acute expression of p75NTR after FPI-TBI and reduction of neurodegeneration with EVT901 in vivo . ( A ) Immunohistochemical expression of p75NTR at 1–2 days in the dentate gyrus of TBI rats using specific anti-p75NTR antibody. ( B ) Representative photomicrographs (original magnification ×10) of Fluoro-Jade® staining of damaged neurons in the granular layer of the dentate gyrus in FPI-TBI rats 1 week post-injury. ( C ) Representative photomicrographs of Fluoro-Jade®-stained sections from the thalamus 14-days post-trauma (ipsilateral versus contralateral to the injury). ( D ) Quantification of Fluoro-Jade® B-positive degenerating cells in the thalamus of FPI-TBI rats after EVT901 treatment. Following Fluoro-Jade staining, the digital images were collected and the damaged cells were quantified using Explora Nova Software. Three mg/kg/day of EVT901 protected neurons in the thalamus after TBI [one-way ANOVA, EVT901 effect, F (3,18) = 8.46, P = 0.001]. # P

    Article Snippet: Anti-rabbit p75NTR Ab (1:1000 dilution; COVANCE) was added and incubated overnight at 4 °C.

    Techniques: Expressing, In Vivo, Immunohistochemistry, Staining, Software

    Mechanism of action through the CRD1 domain of p75NTR: cellular response and induction of neuritogenesis. ( A ) EVT901 inhibits the binding of AP-p75NTR to the receptor p75NTR. Binding experiments on SKN-BE-p75NTR with soluble AP-p75NTR (10 nM) in the presence of increasing concentrations of EVT901. Data are reported as mean % inhibition of binding, representative of three independent experiments. ( B ) Competition of 125 I-NGF binding with EVT901. Binding of 125 I-NGF (0.3 nM) to SKNBE-p75NTR in the presence of increasing concentrations of EVT901. Results are mean % inhibition of specific binding, representative of three independent experiments. ( C ) HEK293T cells transiently co-transfected with the same amount of DNA originating from the p75NTR full-length tagged either in C-terminal with HA (HA- p75NTR) or in N-terminal with Flag (Flag-p75NTR) and then treated or not with the compound EVT901 at 100 nM for 24 h. Oligomerization evaluated using an ELISA assay with flag-coated plates and anti-HA-HRP antibody [two-way ANOVA, EVT901 dose effect, F( 4,94) = 46.85, P

    Journal: Brain

    Article Title: A novel inhibitor of p75-neurotrophin receptor improves functional outcomes in two models of traumatic brain injury

    doi: 10.1093/brain/aww074

    Figure Lengend Snippet: Mechanism of action through the CRD1 domain of p75NTR: cellular response and induction of neuritogenesis. ( A ) EVT901 inhibits the binding of AP-p75NTR to the receptor p75NTR. Binding experiments on SKN-BE-p75NTR with soluble AP-p75NTR (10 nM) in the presence of increasing concentrations of EVT901. Data are reported as mean % inhibition of binding, representative of three independent experiments. ( B ) Competition of 125 I-NGF binding with EVT901. Binding of 125 I-NGF (0.3 nM) to SKNBE-p75NTR in the presence of increasing concentrations of EVT901. Results are mean % inhibition of specific binding, representative of three independent experiments. ( C ) HEK293T cells transiently co-transfected with the same amount of DNA originating from the p75NTR full-length tagged either in C-terminal with HA (HA- p75NTR) or in N-terminal with Flag (Flag-p75NTR) and then treated or not with the compound EVT901 at 100 nM for 24 h. Oligomerization evaluated using an ELISA assay with flag-coated plates and anti-HA-HRP antibody [two-way ANOVA, EVT901 dose effect, F( 4,94) = 46.85, P

    Article Snippet: Anti-rabbit p75NTR Ab (1:1000 dilution; COVANCE) was added and incubated overnight at 4 °C.

    Techniques: Binding Assay, Inhibition, Transfection, Enzyme-linked Immunosorbent Assay

    EVT901 blocks proNGF-mediated caspase activity and p75NTR expression in OPCs and oligodendrocytes in vitro and reduces proNGF-induced cell death. ( A ) Immunostaining showed that treatment with pro-NGF (50 ng/ml) induced caspase-3 activity in OPCs in vitro . ( B and C ) Caspase activity was measured by counting positive cells using the FLICA assay. Pro-NGF (50 ng/ml) increased caspase activity in cultured OPCs and oligodendrocytes. However, EVT901 added to the medium reduced the percentage of caspase-positive cells in a dose-dependent manner for OPCs and oligodendrocytes [one-way ANOVA, F (7,67) = 10.02, P

    Journal: Brain

    Article Title: A novel inhibitor of p75-neurotrophin receptor improves functional outcomes in two models of traumatic brain injury

    doi: 10.1093/brain/aww074

    Figure Lengend Snippet: EVT901 blocks proNGF-mediated caspase activity and p75NTR expression in OPCs and oligodendrocytes in vitro and reduces proNGF-induced cell death. ( A ) Immunostaining showed that treatment with pro-NGF (50 ng/ml) induced caspase-3 activity in OPCs in vitro . ( B and C ) Caspase activity was measured by counting positive cells using the FLICA assay. Pro-NGF (50 ng/ml) increased caspase activity in cultured OPCs and oligodendrocytes. However, EVT901 added to the medium reduced the percentage of caspase-positive cells in a dose-dependent manner for OPCs and oligodendrocytes [one-way ANOVA, F (7,67) = 10.02, P

    Article Snippet: Anti-rabbit p75NTR Ab (1:1000 dilution; COVANCE) was added and incubated overnight at 4 °C.

    Techniques: Activity Assay, Expressing, In Vitro, Immunostaining, Cell Culture