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rabbit anti nnos  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit anti nnos
    Interaction between STRAP and three NOS memebers. A – C , coimmunoprecipitation analysis of STRAP interaction with iNOS ( A ), eNOS ( B ), or nNOS ( C ). HEK293 cells were cotransfected with FLAG-STRAP and iNOS, eNOS, or nNOS expression plasmids. Cell lysates were immunoprecipitated with anti-FLAG antibody or control mouse immunoglobulin G, followed by immunoblotting with anti-iNOS, anti-eNOS, <t>or</t> <t>anti-nNOS</t> antibodies. n = 2. D , detection of S-nitrosylated STRAP (SNO-STRAP) in HEK293 cells overexpressing iNOS, eNOS, or nNOS. Reaction without ascorbate (−Ascorbate) was included as a negative control. n = 3. E , expression levels of iNOS, eNOS, and nNOS in HEK293 cells. eNOS, endothelial nitric oxide synthase; HEK293, human embryonic kidney 293 cell line; iNOS, inducible nitric oxide synthase; NOS, nitric oxide synthase; nNOS, neuronal nitric oxide synthase; SNO, S-nitrosolthiol; STRAP, serine–threonine kinase receptor–associated protein.
    Rabbit Anti Nnos, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 391 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/rabbit+anti+nnos+antibody/eNOS+Rabbit+mAb/pmc12906178-193-31-33
    Average 96 stars, based on 391 article reviews
    rabbit anti nnos - by Bioz Stars, 2026-09
    96/100 stars

    Images

    1) Product Images from "S-nitrosylation of the scaffold protein STRAP enhances oxidative stress–induced apoptosis"

    Article Title: S-nitrosylation of the scaffold protein STRAP enhances oxidative stress–induced apoptosis

    Journal: The Journal of Biological Chemistry

    doi: 10.1016/j.jbc.2026.111141

    Interaction between STRAP and three NOS memebers. A – C , coimmunoprecipitation analysis of STRAP interaction with iNOS ( A ), eNOS ( B ), or nNOS ( C ). HEK293 cells were cotransfected with FLAG-STRAP and iNOS, eNOS, or nNOS expression plasmids. Cell lysates were immunoprecipitated with anti-FLAG antibody or control mouse immunoglobulin G, followed by immunoblotting with anti-iNOS, anti-eNOS, or anti-nNOS antibodies. n = 2. D , detection of S-nitrosylated STRAP (SNO-STRAP) in HEK293 cells overexpressing iNOS, eNOS, or nNOS. Reaction without ascorbate (−Ascorbate) was included as a negative control. n = 3. E , expression levels of iNOS, eNOS, and nNOS in HEK293 cells. eNOS, endothelial nitric oxide synthase; HEK293, human embryonic kidney 293 cell line; iNOS, inducible nitric oxide synthase; NOS, nitric oxide synthase; nNOS, neuronal nitric oxide synthase; SNO, S-nitrosolthiol; STRAP, serine–threonine kinase receptor–associated protein.
    Figure Legend Snippet: Interaction between STRAP and three NOS memebers. A – C , coimmunoprecipitation analysis of STRAP interaction with iNOS ( A ), eNOS ( B ), or nNOS ( C ). HEK293 cells were cotransfected with FLAG-STRAP and iNOS, eNOS, or nNOS expression plasmids. Cell lysates were immunoprecipitated with anti-FLAG antibody or control mouse immunoglobulin G, followed by immunoblotting with anti-iNOS, anti-eNOS, or anti-nNOS antibodies. n = 2. D , detection of S-nitrosylated STRAP (SNO-STRAP) in HEK293 cells overexpressing iNOS, eNOS, or nNOS. Reaction without ascorbate (−Ascorbate) was included as a negative control. n = 3. E , expression levels of iNOS, eNOS, and nNOS in HEK293 cells. eNOS, endothelial nitric oxide synthase; HEK293, human embryonic kidney 293 cell line; iNOS, inducible nitric oxide synthase; NOS, nitric oxide synthase; nNOS, neuronal nitric oxide synthase; SNO, S-nitrosolthiol; STRAP, serine–threonine kinase receptor–associated protein.

    Techniques Used: Expressing, Immunoprecipitation, Control, Western Blot, Negative Control

    Related Articles

    Saline:

    Article Title: Nitric Oxide Mediates Selective Degeneration of Hypothalamic Orexin Neurons through Dysfunction of Protein Disulfide Isomerase
    Article Snippet: .. The blots were washed with Tris-buffered saline containing 0.1% Tween 20 and blocked with Blocking One (Nacalai) at 22–25°C for 1 h. The membrane was incubated with mouse anti-KDEL antibody [for detection of Ig binding protein (BiP), 1:2000; SPA-827, Stressgen, Enzo Life Sciences], rabbit anti-PDI antibody (1:2000; ADI-SPA-890, Enzo Life Sciences), rabbit anti-nNOS antibody (1:1000; Cell Signaling Technology), mouse anti-iNOS antibody (1:1000; 610328, BD Biosciences), and mouse anti-β-actin antibody (1:1000; Sigma-Aldrich) overnight at 4°C. .. After incubation with horseradish peroxidase-conjugated secondary antibodies at 22–25°C for 1 h, bands were detected with ECL Advance Western blotting detection kit (GE Healthcare) on a lumino-imaging analyzer (LAS-3000mini; Fuji Film).

    Blocking Assay:

    Article Title: Nitric Oxide Mediates Selective Degeneration of Hypothalamic Orexin Neurons through Dysfunction of Protein Disulfide Isomerase
    Article Snippet: .. The blots were washed with Tris-buffered saline containing 0.1% Tween 20 and blocked with Blocking One (Nacalai) at 22–25°C for 1 h. The membrane was incubated with mouse anti-KDEL antibody [for detection of Ig binding protein (BiP), 1:2000; SPA-827, Stressgen, Enzo Life Sciences], rabbit anti-PDI antibody (1:2000; ADI-SPA-890, Enzo Life Sciences), rabbit anti-nNOS antibody (1:1000; Cell Signaling Technology), mouse anti-iNOS antibody (1:1000; 610328, BD Biosciences), and mouse anti-β-actin antibody (1:1000; Sigma-Aldrich) overnight at 4°C. .. After incubation with horseradish peroxidase-conjugated secondary antibodies at 22–25°C for 1 h, bands were detected with ECL Advance Western blotting detection kit (GE Healthcare) on a lumino-imaging analyzer (LAS-3000mini; Fuji Film).

    Membrane:

    Article Title: Nitric Oxide Mediates Selective Degeneration of Hypothalamic Orexin Neurons through Dysfunction of Protein Disulfide Isomerase
    Article Snippet: .. The blots were washed with Tris-buffered saline containing 0.1% Tween 20 and blocked with Blocking One (Nacalai) at 22–25°C for 1 h. The membrane was incubated with mouse anti-KDEL antibody [for detection of Ig binding protein (BiP), 1:2000; SPA-827, Stressgen, Enzo Life Sciences], rabbit anti-PDI antibody (1:2000; ADI-SPA-890, Enzo Life Sciences), rabbit anti-nNOS antibody (1:1000; Cell Signaling Technology), mouse anti-iNOS antibody (1:1000; 610328, BD Biosciences), and mouse anti-β-actin antibody (1:1000; Sigma-Aldrich) overnight at 4°C. .. After incubation with horseradish peroxidase-conjugated secondary antibodies at 22–25°C for 1 h, bands were detected with ECL Advance Western blotting detection kit (GE Healthcare) on a lumino-imaging analyzer (LAS-3000mini; Fuji Film).

    Incubation:

    Article Title: Nitric Oxide Mediates Selective Degeneration of Hypothalamic Orexin Neurons through Dysfunction of Protein Disulfide Isomerase
    Article Snippet: .. The blots were washed with Tris-buffered saline containing 0.1% Tween 20 and blocked with Blocking One (Nacalai) at 22–25°C for 1 h. The membrane was incubated with mouse anti-KDEL antibody [for detection of Ig binding protein (BiP), 1:2000; SPA-827, Stressgen, Enzo Life Sciences], rabbit anti-PDI antibody (1:2000; ADI-SPA-890, Enzo Life Sciences), rabbit anti-nNOS antibody (1:1000; Cell Signaling Technology), mouse anti-iNOS antibody (1:1000; 610328, BD Biosciences), and mouse anti-β-actin antibody (1:1000; Sigma-Aldrich) overnight at 4°C. .. After incubation with horseradish peroxidase-conjugated secondary antibodies at 22–25°C for 1 h, bands were detected with ECL Advance Western blotting detection kit (GE Healthcare) on a lumino-imaging analyzer (LAS-3000mini; Fuji Film).

    Article Title: Gastric nNOS reduction accompanied by natriuretic peptides signaling pathway upregulation in diabetic mice
    Article Snippet: .. To reduce non-specific antibody binding, they were preincubated in 5% bovine serum albumin (Sigma) for 1 h at room temperature before incubation with the rabbit anti-nNOS antibody (1:1000; Cell Signaling Technology, Danvers, MA, United States) and mouse anti-HuC/HuD antibody (A-21271, Abcam, Burlingame, CA, United States). .. To achieve greater penetration during labeling, incubation solutions with the primary antibody were mixed with Triton-X 100 (0.5%; Sigma).

    Article Title: Mitogen-activated protein kinases regulate expression of neuronal nitric oxide synthase and neurite outgrowth via non-classical retinoic acid receptor signaling in human neuroblastoma SH-SY5Y cells.
    Article Snippet: The membrane was washed with Tris-buffered saline/Tween 20 and blocked either with Blocking One (#03953-95, Nacalai Tesque, Kyoto, Japan) for nNOS, DAX1 and b-actin, or with Blocking One-P (#05999-84, Nacalai Tesque) for c-Jun N-terminal kinase (JNK) and p38 MAPK, at 22e25 C for 1 h. The membrane was incubated with rabbit anti-phospho-JNK antibody [phospho-SAPK/ JNK(Thr183/Tyr185), 1:1000; #9251, Cell Signaling Tech., Danvers, MA, USA], rabbit anti-JNK antibody (1:1000; #9252, Cell Signaling Tech.), rabbit anti-phospho-p38 MAPK antibody [phospho-p38 MAP kinase (Thr180/Tyr182), 1:1000; #9215S, Cell Signaling Tech. .. ], rabbit anti-p38 MAPK antibody (1:1000; #9212, Cell Signaling Tech.), rabbit anti-nNOS antibody (1:1,000, #4234, Cell Signaling Tech.), rabbit anti-DAX1 antibody (1:1000; #sc-841, Santa Cruz Biotechnology, Inc.) andmouse anti-b-actin antibody (1:1,000, #A5441, SigmaeAldrich) overnight at 4 C. After incubation with horseradish peroxidase-conjugated secondary antibodies at 22e25 C for 1 h, bands were detected with ECL Select Western blotting detection kit (#RPN2235, Amersham Biosciences, Piscataway, NJ, USA) on a lumino-imaging analyzer (LAS-3000mini, Fuji Film, Tokyo, Japan). .. Real-time PCR was performed with SYBR® Premix Ex TaqTM (#RR420S, TaKaRa) on a Chromo4TM real-time PCR analysis system (Bio-Rad, Tokyo, Japan).

    Article Title: Diabetes-induced damage of gastric nitric oxide neurons mediated by P2X7R in diabetic mice.
    Article Snippet: It is generally considered that enteric neuropathy is one of the causative factors in diabetic gastroparesis.. Our previous study demonstrated that there is a loss of NOS neurons in diabetic mice.. However, the underlying mechanism remains unclear.

    Binding Assay:

    Article Title: Nitric Oxide Mediates Selective Degeneration of Hypothalamic Orexin Neurons through Dysfunction of Protein Disulfide Isomerase
    Article Snippet: .. The blots were washed with Tris-buffered saline containing 0.1% Tween 20 and blocked with Blocking One (Nacalai) at 22–25°C for 1 h. The membrane was incubated with mouse anti-KDEL antibody [for detection of Ig binding protein (BiP), 1:2000; SPA-827, Stressgen, Enzo Life Sciences], rabbit anti-PDI antibody (1:2000; ADI-SPA-890, Enzo Life Sciences), rabbit anti-nNOS antibody (1:1000; Cell Signaling Technology), mouse anti-iNOS antibody (1:1000; 610328, BD Biosciences), and mouse anti-β-actin antibody (1:1000; Sigma-Aldrich) overnight at 4°C. .. After incubation with horseradish peroxidase-conjugated secondary antibodies at 22–25°C for 1 h, bands were detected with ECL Advance Western blotting detection kit (GE Healthcare) on a lumino-imaging analyzer (LAS-3000mini; Fuji Film).

    Article Title: Gastric nNOS reduction accompanied by natriuretic peptides signaling pathway upregulation in diabetic mice
    Article Snippet: .. To reduce non-specific antibody binding, they were preincubated in 5% bovine serum albumin (Sigma) for 1 h at room temperature before incubation with the rabbit anti-nNOS antibody (1:1000; Cell Signaling Technology, Danvers, MA, United States) and mouse anti-HuC/HuD antibody (A-21271, Abcam, Burlingame, CA, United States). .. To achieve greater penetration during labeling, incubation solutions with the primary antibody were mixed with Triton-X 100 (0.5%; Sigma).

    Western Blot:

    Article Title: Mitogen-activated protein kinases regulate expression of neuronal nitric oxide synthase and neurite outgrowth via non-classical retinoic acid receptor signaling in human neuroblastoma SH-SY5Y cells.
    Article Snippet: The membrane was washed with Tris-buffered saline/Tween 20 and blocked either with Blocking One (#03953-95, Nacalai Tesque, Kyoto, Japan) for nNOS, DAX1 and b-actin, or with Blocking One-P (#05999-84, Nacalai Tesque) for c-Jun N-terminal kinase (JNK) and p38 MAPK, at 22e25 C for 1 h. The membrane was incubated with rabbit anti-phospho-JNK antibody [phospho-SAPK/ JNK(Thr183/Tyr185), 1:1000; #9251, Cell Signaling Tech., Danvers, MA, USA], rabbit anti-JNK antibody (1:1000; #9252, Cell Signaling Tech.), rabbit anti-phospho-p38 MAPK antibody [phospho-p38 MAP kinase (Thr180/Tyr182), 1:1000; #9215S, Cell Signaling Tech. .. ], rabbit anti-p38 MAPK antibody (1:1000; #9212, Cell Signaling Tech.), rabbit anti-nNOS antibody (1:1,000, #4234, Cell Signaling Tech.), rabbit anti-DAX1 antibody (1:1000; #sc-841, Santa Cruz Biotechnology, Inc.) andmouse anti-b-actin antibody (1:1,000, #A5441, SigmaeAldrich) overnight at 4 C. After incubation with horseradish peroxidase-conjugated secondary antibodies at 22e25 C for 1 h, bands were detected with ECL Select Western blotting detection kit (#RPN2235, Amersham Biosciences, Piscataway, NJ, USA) on a lumino-imaging analyzer (LAS-3000mini, Fuji Film, Tokyo, Japan). .. Real-time PCR was performed with SYBR® Premix Ex TaqTM (#RR420S, TaKaRa) on a Chromo4TM real-time PCR analysis system (Bio-Rad, Tokyo, Japan).



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    Interaction between STRAP and three NOS memebers. A – C , coimmunoprecipitation analysis of STRAP interaction with iNOS ( A ), eNOS ( B ), or nNOS ( C ). HEK293 cells were cotransfected with FLAG-STRAP and iNOS, eNOS, or nNOS expression plasmids. Cell lysates were immunoprecipitated with anti-FLAG antibody or control mouse immunoglobulin G, followed by immunoblotting with anti-iNOS, anti-eNOS, <t>or</t> <t>anti-nNOS</t> antibodies. n = 2. D , detection of S-nitrosylated STRAP (SNO-STRAP) in HEK293 cells overexpressing iNOS, eNOS, or nNOS. Reaction without ascorbate (−Ascorbate) was included as a negative control. n = 3. E , expression levels of iNOS, eNOS, and nNOS in HEK293 cells. eNOS, endothelial nitric oxide synthase; HEK293, human embryonic kidney 293 cell line; iNOS, inducible nitric oxide synthase; NOS, nitric oxide synthase; nNOS, neuronal nitric oxide synthase; SNO, S-nitrosolthiol; STRAP, serine–threonine kinase receptor–associated protein.
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    Enhanced intercellular communication drives neuroinflammatory responses following cardiac arrest. (A) Heatmap illustrating the numbers of inferred interactions between different cell types based on LR pairs. (B) Chord diagram of the top 30 LR pairs between cell types. (C, D) Network diagrams of the changes in the number of intercellular communications between the Sham and ROSC6h groups (C) and the Sham and ROSC24h groups (D). Red represents an increase, and blue represents a decrease, in the number of interactions. (E) Visualization of enhanced LR pairs involved in microglia–neutrophil communication after cardiac arrest in the ROSC6h and ROSC24h groups vs. the Sham group. Each dot represents an LR pair, with its position indicating the group. Dot size corresponds to the P value (larger dots indicate lower P values), while the color represents the interaction probability, with the gradient ranging from purple (low probability) to yellow (high probability). (F) Secretion of resistin into the supernatant of the neutrophil–microglia co-culture system over time. (G) Comparison of resistin secretion, with and without OGD/R and the presence of neutrophils. (H) Immunofluorescence analysis and quantification of <t>iNOS</t> expression in microglia. The left panel shows representative images of iNOS (red), IBA1 (green, microglia marker), and DAPI (blue, nuclei), with merged images in the bottom row. The bar graph shows quantification of relative iNOS fluorescence intensity. Statistical significance determined by one-way analysis of variance followed by Tukey’s post hoc test; **** P < 0.0001. Scale bars: 20 μm. IBA1: Ionized calcium binding adaptor molecule 1; iNOS: inducible nitric oxide <t>synthase;</t> LR: ligand receptor; ns: not significant; OGD/R: oxygen-glucose deprivation/reperfusion; ROSC: return of spontaneous circulation.
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    Results of immunofluorescence assays. a Immunofluorescence photographs of synaptophysin (red) and the cell nucleus (blue). b Immunofluorescence photographs of Brain Derived Neurotrophic Factor (red) and the cell nucleus (blue). c Immunofluorescence photographs <t>of</t> <t>α-synuclein</t> (red) and the cell nucleus (blue). d Immunofluorescence photographs of neuronal Nitric Oxide Synthase (green) and the cell nucleus (blue). e Immunofluorescence photographs of β-actin (red) and the cell nucleus (blue). f CBL increases synaptophysin in prefrontal cortex and hippocampus. g CBL increases the presence of Brain Derived Neurotrophic Factor in the prefrontal cortex, CA1 hippocampus, and basolateral amygdala. h α-synuclein is not affected by CBL treatment. i CBL treatment did not affect <t>nNOS</t> density. j CBL induces an increase in β-actin in each analyzed region. (PFC: Prefrontal cortex; DG: Dentate gyrus; BLA: basolateral amygdala; Syn; Synaptophysin; BDNF; Brain derived neurotrophic factor; nNOS: neuronal oxide nitric synthase)
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    Image Search Results


    Interaction between STRAP and three NOS memebers. A – C , coimmunoprecipitation analysis of STRAP interaction with iNOS ( A ), eNOS ( B ), or nNOS ( C ). HEK293 cells were cotransfected with FLAG-STRAP and iNOS, eNOS, or nNOS expression plasmids. Cell lysates were immunoprecipitated with anti-FLAG antibody or control mouse immunoglobulin G, followed by immunoblotting with anti-iNOS, anti-eNOS, or anti-nNOS antibodies. n = 2. D , detection of S-nitrosylated STRAP (SNO-STRAP) in HEK293 cells overexpressing iNOS, eNOS, or nNOS. Reaction without ascorbate (−Ascorbate) was included as a negative control. n = 3. E , expression levels of iNOS, eNOS, and nNOS in HEK293 cells. eNOS, endothelial nitric oxide synthase; HEK293, human embryonic kidney 293 cell line; iNOS, inducible nitric oxide synthase; NOS, nitric oxide synthase; nNOS, neuronal nitric oxide synthase; SNO, S-nitrosolthiol; STRAP, serine–threonine kinase receptor–associated protein.

    Journal: The Journal of Biological Chemistry

    Article Title: S-nitrosylation of the scaffold protein STRAP enhances oxidative stress–induced apoptosis

    doi: 10.1016/j.jbc.2026.111141

    Figure Lengend Snippet: Interaction between STRAP and three NOS memebers. A – C , coimmunoprecipitation analysis of STRAP interaction with iNOS ( A ), eNOS ( B ), or nNOS ( C ). HEK293 cells were cotransfected with FLAG-STRAP and iNOS, eNOS, or nNOS expression plasmids. Cell lysates were immunoprecipitated with anti-FLAG antibody or control mouse immunoglobulin G, followed by immunoblotting with anti-iNOS, anti-eNOS, or anti-nNOS antibodies. n = 2. D , detection of S-nitrosylated STRAP (SNO-STRAP) in HEK293 cells overexpressing iNOS, eNOS, or nNOS. Reaction without ascorbate (−Ascorbate) was included as a negative control. n = 3. E , expression levels of iNOS, eNOS, and nNOS in HEK293 cells. eNOS, endothelial nitric oxide synthase; HEK293, human embryonic kidney 293 cell line; iNOS, inducible nitric oxide synthase; NOS, nitric oxide synthase; nNOS, neuronal nitric oxide synthase; SNO, S-nitrosolthiol; STRAP, serine–threonine kinase receptor–associated protein.

    Article Snippet: Eluted proteins were separated by SDS-PAGE on 4% to 20% Criterion Precast Midi Protein Gels (Bio-Rad), transferred to PVDF membranes, and probed with the following antibodies: rabbit anti-eNOS (Cell Signaling, 32027S), rabbit anti-nNOS (Cell Signaling, 4231S), rabbit anti-iNOS (Santa Cruz, sc-8310), and rabbit anti-myc (Cell Signaling, 2278S).

    Techniques: Expressing, Immunoprecipitation, Control, Western Blot, Negative Control

    Enhanced intercellular communication drives neuroinflammatory responses following cardiac arrest. (A) Heatmap illustrating the numbers of inferred interactions between different cell types based on LR pairs. (B) Chord diagram of the top 30 LR pairs between cell types. (C, D) Network diagrams of the changes in the number of intercellular communications between the Sham and ROSC6h groups (C) and the Sham and ROSC24h groups (D). Red represents an increase, and blue represents a decrease, in the number of interactions. (E) Visualization of enhanced LR pairs involved in microglia–neutrophil communication after cardiac arrest in the ROSC6h and ROSC24h groups vs. the Sham group. Each dot represents an LR pair, with its position indicating the group. Dot size corresponds to the P value (larger dots indicate lower P values), while the color represents the interaction probability, with the gradient ranging from purple (low probability) to yellow (high probability). (F) Secretion of resistin into the supernatant of the neutrophil–microglia co-culture system over time. (G) Comparison of resistin secretion, with and without OGD/R and the presence of neutrophils. (H) Immunofluorescence analysis and quantification of iNOS expression in microglia. The left panel shows representative images of iNOS (red), IBA1 (green, microglia marker), and DAPI (blue, nuclei), with merged images in the bottom row. The bar graph shows quantification of relative iNOS fluorescence intensity. Statistical significance determined by one-way analysis of variance followed by Tukey’s post hoc test; **** P < 0.0001. Scale bars: 20 μm. IBA1: Ionized calcium binding adaptor molecule 1; iNOS: inducible nitric oxide synthase; LR: ligand receptor; ns: not significant; OGD/R: oxygen-glucose deprivation/reperfusion; ROSC: return of spontaneous circulation.

    Journal: Neural Regeneration Research

    Article Title: Blood–brain barrier disruption and neuroinflammation in the hippocampus of a cardiac arrest porcine model: Single-cell RNA sequencing analysis

    doi: 10.4103/NRR.NRR-D-24-01269

    Figure Lengend Snippet: Enhanced intercellular communication drives neuroinflammatory responses following cardiac arrest. (A) Heatmap illustrating the numbers of inferred interactions between different cell types based on LR pairs. (B) Chord diagram of the top 30 LR pairs between cell types. (C, D) Network diagrams of the changes in the number of intercellular communications between the Sham and ROSC6h groups (C) and the Sham and ROSC24h groups (D). Red represents an increase, and blue represents a decrease, in the number of interactions. (E) Visualization of enhanced LR pairs involved in microglia–neutrophil communication after cardiac arrest in the ROSC6h and ROSC24h groups vs. the Sham group. Each dot represents an LR pair, with its position indicating the group. Dot size corresponds to the P value (larger dots indicate lower P values), while the color represents the interaction probability, with the gradient ranging from purple (low probability) to yellow (high probability). (F) Secretion of resistin into the supernatant of the neutrophil–microglia co-culture system over time. (G) Comparison of resistin secretion, with and without OGD/R and the presence of neutrophils. (H) Immunofluorescence analysis and quantification of iNOS expression in microglia. The left panel shows representative images of iNOS (red), IBA1 (green, microglia marker), and DAPI (blue, nuclei), with merged images in the bottom row. The bar graph shows quantification of relative iNOS fluorescence intensity. Statistical significance determined by one-way analysis of variance followed by Tukey’s post hoc test; **** P < 0.0001. Scale bars: 20 μm. IBA1: Ionized calcium binding adaptor molecule 1; iNOS: inducible nitric oxide synthase; LR: ligand receptor; ns: not significant; OGD/R: oxygen-glucose deprivation/reperfusion; ROSC: return of spontaneous circulation.

    Article Snippet: They were then incubated with the following primary antibodies at 4°C overnight: rabbit anti-ionized calcium binding adaptor molecule 1 (IBA1) (1:200 dilution; Abcam, Cambridge, UK, Cat# ab178846, RRID: AB_2636859); mouse anti-S100A8 (1:200; Proteintech, Wuhan, China, Cat# 66853-1-Ig, RRID: AB_2882193); rabbit anti-myelin basic protein (MBP) (1:50, Cell Signaling Technology, Danvers, MA, USA; Cat# 78896, RRID: AB_2799920); rabbit anti-myeloperoxidase (MPO) (1:100, Abcam, Cat# ab208670, RRID: AB_2864724); mouse anti-IBA1 (1:200, Santa Cruz Biotechnology, Santa Cruz, CA, USA, Cat# sc-32725, RRID: AB_667733); and rabbit anti-inducible nitric oxide synthase (iNOS) (1:200, Proteintech, Cat# 18985-1-AP, RRID: AB_2782960).

    Techniques: Co-Culture Assay, Comparison, Immunofluorescence, Expressing, Marker, Fluorescence, Binding Assay

    Results of immunofluorescence assays. a Immunofluorescence photographs of synaptophysin (red) and the cell nucleus (blue). b Immunofluorescence photographs of Brain Derived Neurotrophic Factor (red) and the cell nucleus (blue). c Immunofluorescence photographs of α-synuclein (red) and the cell nucleus (blue). d Immunofluorescence photographs of neuronal Nitric Oxide Synthase (green) and the cell nucleus (blue). e Immunofluorescence photographs of β-actin (red) and the cell nucleus (blue). f CBL increases synaptophysin in prefrontal cortex and hippocampus. g CBL increases the presence of Brain Derived Neurotrophic Factor in the prefrontal cortex, CA1 hippocampus, and basolateral amygdala. h α-synuclein is not affected by CBL treatment. i CBL treatment did not affect nNOS density. j CBL induces an increase in β-actin in each analyzed region. (PFC: Prefrontal cortex; DG: Dentate gyrus; BLA: basolateral amygdala; Syn; Synaptophysin; BDNF; Brain derived neurotrophic factor; nNOS: neuronal oxide nitric synthase)

    Journal: Neurochemical Research

    Article Title: Cerebrolysin Ameliorates Age-Induced Dendritic Spine Degeneration and Memory Decline in C57BL6 Mice

    doi: 10.1007/s11064-025-04627-0

    Figure Lengend Snippet: Results of immunofluorescence assays. a Immunofluorescence photographs of synaptophysin (red) and the cell nucleus (blue). b Immunofluorescence photographs of Brain Derived Neurotrophic Factor (red) and the cell nucleus (blue). c Immunofluorescence photographs of α-synuclein (red) and the cell nucleus (blue). d Immunofluorescence photographs of neuronal Nitric Oxide Synthase (green) and the cell nucleus (blue). e Immunofluorescence photographs of β-actin (red) and the cell nucleus (blue). f CBL increases synaptophysin in prefrontal cortex and hippocampus. g CBL increases the presence of Brain Derived Neurotrophic Factor in the prefrontal cortex, CA1 hippocampus, and basolateral amygdala. h α-synuclein is not affected by CBL treatment. i CBL treatment did not affect nNOS density. j CBL induces an increase in β-actin in each analyzed region. (PFC: Prefrontal cortex; DG: Dentate gyrus; BLA: basolateral amygdala; Syn; Synaptophysin; BDNF; Brain derived neurotrophic factor; nNOS: neuronal oxide nitric synthase)

    Article Snippet: Slides were incubated for 24 h at 4 °C with the primary antibody to Syn (Cell Signaling #36406, Clone: D8F6H, 1:100), BDNF (Abcam, clone: 3C11, 5 μg/mL), α-synuclein (Cell Signaling #2647, clone: Syn204, 1:1000), nNOS (Cell Signaling #4231, Clone: C7D7, 1:200), and β-actin (Cell Signaling #3700, clone: 8H10D10, 1:3000).

    Techniques: Immunofluorescence, Derivative Assay