rabbit anti nav1 7  (Alomone Labs)


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    Alomone Labs rabbit anti nav1 7
    Rabbit Anti Nav1 7, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 95 stars, based on 1 article reviews
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    Alomone Labs p2ry12
    Interaction of <t>P2RY12-positive</t> microglia with different types of Aβ-positive plaques in pathologically-staged cases. ( A – F ). P2RY12-positive microglia (green) interacting with Aβ-positive diffuse-type plaques (red) ( A , C , E ), in LPND ( A ), HPND ( C ), and AD ( E ), but not with mature-type cored plaques ( B -LPND) ( D -HPND) ( F -AD). Scale bars represent 50 μm.
    P2ry12, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 86 stars, based on 1 article reviews
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    95
    Alomone Labs rabbit anti hcn1
    Quantitative polymerase chain reaction (qPCR) ΔCt analysis for hyperpolarization-activated cyclic nucleotide-gated channel (HCN channel) subunits in proprioceptive DRG cells. (A) Box and whisker plot show group data comparing ΔCt for GFP expression in GFP+ (green) and GFP− (blue) samples (normalized to β actin expression). The low GFP ΔCt in GFP+, but not GFP−, confirm high expression in this sample and the reliability of this sampling approach. (B) Box whisker plots plot shows group data comparing ΔCt values for <t>HCN1–4</t> in GFP+ (green) and GFP− (blue) samples. The expression of all HCN subunits was significantly different between GFP+ and GFP− groups (where * p
    Rabbit Anti Hcn1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Alomone Labs kt5823
    Inhibition of PKG prevented the proliferation of SVZ cells stimulated by PDE5 inhibitors. Cell proliferation following treatment with 1 μ M <t>KT5823</t> and 1 μ M T0156 (a, d), 1 μ M sildenafil (b, e), or 10 μ M zaprinast (c, f) was assessed by incorporation of EdU and evaluated by flow cytometry, following 6 h or 24 h of treatment. Data are expressed as means ± SEM of at least 4 independent experiments. One-way ANOVA (Bonferroni's post-test), * P
    Kt5823, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs mature bdnf
    Line-blot for qualitative analysis of <t>anti-BDNF</t> antibodies specificity. ( A ) The antibodies from each ELISA kit were tested for specificity against pro-BDNF or mature BDNF. The BDNF standards blotted were commercial pro-BDNF (Alomone; 10 pg/lane), mature BDNF (1 and 2 from Alomone and Sigma, respectively; both 1000 pg/lane) and the standard BDNF protein included in each kit (Aviscera-Bioscience and Biosensis: 10 pg/lane; Millipore-ChemiKine TM , Millipore-Milliplex ® - and R D <t>System-Quantikine</t> ® : 100 pg/lane; Promega-Emax ® : 1000 pg/lane). BSA (1000 pg/lane) was used as a negative control. The mouse monoclonal anti-BDNF antibody, (1:1000; Sigma) was tested as a control. ( B ) Central region of the same blot shown in A, from an overexposed film to better visualize the reactivity against pro-BDNF. ( C ) Reactivity of antibodies from Biosensis, Promega-Emax ® pAb and Sigma on a dot blot in which the same quantity of pro-BNDF and mature BDNF were spotted (100 pg each). Each antibody from the ELISA kits was used at the dilution suggested by the manufacturer’s instructions. mAb: Promega-Emax ® monoclonal capture antibody for plate coating. pAb: Promega-Emax ® polyclonal detection antibody.
    Mature Bdnf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Interaction of P2RY12-positive microglia with different types of Aβ-positive plaques in pathologically-staged cases. ( A – F ). P2RY12-positive microglia (green) interacting with Aβ-positive diffuse-type plaques (red) ( A , C , E ), in LPND ( A ), HPND ( C ), and AD ( E ), but not with mature-type cored plaques ( B -LPND) ( D -HPND) ( F -AD). Scale bars represent 50 μm.

    Journal: International Journal of Molecular Sciences

    Article Title: Patterns of Expression of Purinergic Receptor P2RY12, a Putative Marker for Non-Activated Microglia, in Aged and Alzheimer’s Disease Brains

    doi: 10.3390/ijms21020678

    Figure Lengend Snippet: Interaction of P2RY12-positive microglia with different types of Aβ-positive plaques in pathologically-staged cases. ( A – F ). P2RY12-positive microglia (green) interacting with Aβ-positive diffuse-type plaques (red) ( A , C , E ), in LPND ( A ), HPND ( C ), and AD ( E ), but not with mature-type cored plaques ( B -LPND) ( D -HPND) ( F -AD). Scale bars represent 50 μm.

    Article Snippet: In addition, comparisons of immunostaining patterns were carried using an independent antibody to P2RY12 (1:250, Alomone Labs), prepared against an 18-amino acid peptide sequence that did not overlap with the immunizing sequence of the Novus antibody.

    Techniques:

    Confocal microscopy of P2RY12 and progranulin-positive microglia in pathologically staged cases. ( A – C ). Low magnification merged images of P2RY12 (green), progranulin (PGRN) (red) and DAPI (blue) in MTG of LPND ( A ), HPND ( B ) and AD cases ( C ) to show the distribution of P2RY12 and PGRN immunoreactivity through cortical layers. Scale bar represents 50 μm. ( D – F ). Higher magnification images of PGRN (red) in MTG of LPND ( D ), HPND ( E ) and AD cases ( F ) to show the distribution of immunoreactivity. Similar amounts of PGRN immunoreactivity was present in each disease group. Scale bar represents 50 μm. ( G – I ). Merged images of P2RY12with PGRNimages shown in ( D – F ) with DAPI (blue) showing expression in same cells (yellow arrows). Scale bar represents 25 μm.

    Journal: International Journal of Molecular Sciences

    Article Title: Patterns of Expression of Purinergic Receptor P2RY12, a Putative Marker for Non-Activated Microglia, in Aged and Alzheimer’s Disease Brains

    doi: 10.3390/ijms21020678

    Figure Lengend Snippet: Confocal microscopy of P2RY12 and progranulin-positive microglia in pathologically staged cases. ( A – C ). Low magnification merged images of P2RY12 (green), progranulin (PGRN) (red) and DAPI (blue) in MTG of LPND ( A ), HPND ( B ) and AD cases ( C ) to show the distribution of P2RY12 and PGRN immunoreactivity through cortical layers. Scale bar represents 50 μm. ( D – F ). Higher magnification images of PGRN (red) in MTG of LPND ( D ), HPND ( E ) and AD cases ( F ) to show the distribution of immunoreactivity. Similar amounts of PGRN immunoreactivity was present in each disease group. Scale bar represents 50 μm. ( G – I ). Merged images of P2RY12with PGRNimages shown in ( D – F ) with DAPI (blue) showing expression in same cells (yellow arrows). Scale bar represents 25 μm.

    Article Snippet: In addition, comparisons of immunostaining patterns were carried using an independent antibody to P2RY12 (1:250, Alomone Labs), prepared against an 18-amino acid peptide sequence that did not overlap with the immunizing sequence of the Novus antibody.

    Techniques: Confocal Microscopy, Expressing

    Quantitative biochemical measurements of P2RY12 protein and mRNA in human brains. ( A–C ). Western blot measurements of P2RY12 levels in MTG samples from LP, HP and AD brains. ( A ). Representative western blot image of P2RY12 polypeptide of MTG protein extracts identified with Novus antibody. Blots were normalized for levels of β actin. ( B ). Scatter plot showing individual P2RY12 expression levels. Significant decrease in protein levels of 58 kDa full-length P2RY12 band in AD (green shapes) compared to LPND (black) and HPND (red) cases. Chart indicates mean + Standard error of mean (SEM). Statistical analysis by one-way ANOVA with Tukey post-hoc test (F 2,26 = 11.54, p

    Journal: International Journal of Molecular Sciences

    Article Title: Patterns of Expression of Purinergic Receptor P2RY12, a Putative Marker for Non-Activated Microglia, in Aged and Alzheimer’s Disease Brains

    doi: 10.3390/ijms21020678

    Figure Lengend Snippet: Quantitative biochemical measurements of P2RY12 protein and mRNA in human brains. ( A–C ). Western blot measurements of P2RY12 levels in MTG samples from LP, HP and AD brains. ( A ). Representative western blot image of P2RY12 polypeptide of MTG protein extracts identified with Novus antibody. Blots were normalized for levels of β actin. ( B ). Scatter plot showing individual P2RY12 expression levels. Significant decrease in protein levels of 58 kDa full-length P2RY12 band in AD (green shapes) compared to LPND (black) and HPND (red) cases. Chart indicates mean + Standard error of mean (SEM). Statistical analysis by one-way ANOVA with Tukey post-hoc test (F 2,26 = 11.54, p

    Article Snippet: In addition, comparisons of immunostaining patterns were carried using an independent antibody to P2RY12 (1:250, Alomone Labs), prepared against an 18-amino acid peptide sequence that did not overlap with the immunizing sequence of the Novus antibody.

    Techniques: Western Blot, Expressing

    Proposed scheme of arrangement of different P2RY12-expressing microglia around Aβ plaques. Suggested scheme to describe localized areas of microglial inflammation around plaques. Zone 1: microglia interacting with mature plaques (HLA-DR high, P2RY12 negative) producing proinflammatory cytokines. Zone 2: Area adjacent to plaque with low or negative P2RY12 positive microglia. Zone 3. P2RY12 high expression in surrounding area defining the boundary between proinflammatory area (Zone 1 and 2) and non-affected area (Zone 3 and beyond). As described in this report, exceptions to this scheme were observed.

    Journal: International Journal of Molecular Sciences

    Article Title: Patterns of Expression of Purinergic Receptor P2RY12, a Putative Marker for Non-Activated Microglia, in Aged and Alzheimer’s Disease Brains

    doi: 10.3390/ijms21020678

    Figure Lengend Snippet: Proposed scheme of arrangement of different P2RY12-expressing microglia around Aβ plaques. Suggested scheme to describe localized areas of microglial inflammation around plaques. Zone 1: microglia interacting with mature plaques (HLA-DR high, P2RY12 negative) producing proinflammatory cytokines. Zone 2: Area adjacent to plaque with low or negative P2RY12 positive microglia. Zone 3. P2RY12 high expression in surrounding area defining the boundary between proinflammatory area (Zone 1 and 2) and non-affected area (Zone 3 and beyond). As described in this report, exceptions to this scheme were observed.

    Article Snippet: In addition, comparisons of immunostaining patterns were carried using an independent antibody to P2RY12 (1:250, Alomone Labs), prepared against an 18-amino acid peptide sequence that did not overlap with the immunizing sequence of the Novus antibody.

    Techniques: Expressing

    Quantitative measurements of P2RY12 immunoreactive structures in human brain tissue sections. ( A , B ). Measurements of area occupied of P2RY12 immunoreactivity in LPND ( n = 11) (black), HPND ( n = 11) (red) and AD ( n = 12) (green) cases. ( A ). Sections were imaged at 4x magnification (three random areas/cases) and area occupied in thresholded images measured using Image J software. Results show significant decrease between HPND and AD cases. Statistical analysis by one-way ANOVA with Tukey post-hoc test (F 2,29 = 3.903, p

    Journal: International Journal of Molecular Sciences

    Article Title: Patterns of Expression of Purinergic Receptor P2RY12, a Putative Marker for Non-Activated Microglia, in Aged and Alzheimer’s Disease Brains

    doi: 10.3390/ijms21020678

    Figure Lengend Snippet: Quantitative measurements of P2RY12 immunoreactive structures in human brain tissue sections. ( A , B ). Measurements of area occupied of P2RY12 immunoreactivity in LPND ( n = 11) (black), HPND ( n = 11) (red) and AD ( n = 12) (green) cases. ( A ). Sections were imaged at 4x magnification (three random areas/cases) and area occupied in thresholded images measured using Image J software. Results show significant decrease between HPND and AD cases. Statistical analysis by one-way ANOVA with Tukey post-hoc test (F 2,29 = 3.903, p

    Article Snippet: In addition, comparisons of immunostaining patterns were carried using an independent antibody to P2RY12 (1:250, Alomone Labs), prepared against an 18-amino acid peptide sequence that did not overlap with the immunizing sequence of the Novus antibody.

    Techniques: Software

    Distribution of P2RY12 microglia within cortical layers in relation to amyloid beta plaques in pathologically staged samples ( A – C ). Lower magnification photomicrographs showing the changes in P2RY12 microglia distribution compared to Aβ plaques within cortical layers of MTG. Sections from low plaque, high plaque and AD cases stained for P2RY12 (purple) and Aβ (brown). Sections were counterstained with neutral red to identity cellular morphology. Scale bars represent 200 μm. ( D – F ). Higher magnification photomicrographs of the areas in panels ( A – C ) indicated by frames. Scale bars represent 50 μm.

    Journal: International Journal of Molecular Sciences

    Article Title: Patterns of Expression of Purinergic Receptor P2RY12, a Putative Marker for Non-Activated Microglia, in Aged and Alzheimer’s Disease Brains

    doi: 10.3390/ijms21020678

    Figure Lengend Snippet: Distribution of P2RY12 microglia within cortical layers in relation to amyloid beta plaques in pathologically staged samples ( A – C ). Lower magnification photomicrographs showing the changes in P2RY12 microglia distribution compared to Aβ plaques within cortical layers of MTG. Sections from low plaque, high plaque and AD cases stained for P2RY12 (purple) and Aβ (brown). Sections were counterstained with neutral red to identity cellular morphology. Scale bars represent 200 μm. ( D – F ). Higher magnification photomicrographs of the areas in panels ( A – C ) indicated by frames. Scale bars represent 50 μm.

    Article Snippet: In addition, comparisons of immunostaining patterns were carried using an independent antibody to P2RY12 (1:250, Alomone Labs), prepared against an 18-amino acid peptide sequence that did not overlap with the immunizing sequence of the Novus antibody.

    Techniques: Staining

    Features of P2RY12-immunoreactive microglia. ( A , B ). Morphology of P2RY12-immunoreactive microglia (purple) in a low plaque non-demented (LPND) case ( A ) and AD case ( B ). Sections of middle temporal gyrus (MTG) were single-stained with antibody to P2RY12. Red arrows in panel B illustrate the lack of P2RY12 immunoreactive cells in an area occupied by plaque. ( C , D ). P2RY12-immunoreactive microglia (purple) are a feature in hippocampus sections from non-demented (ND) case ( H ) and Alzheimer’s disease (AD) case. Section shows staining in CA2 region of hippocampus. Continued presence of P2RY12-positive microglia in AD hippocampus ( D ) was noticeable. ( E , F ). Double-staining of section of ND and AD case with P2RY12 (purple) and HLA-DR (brown) showed limited overlap. HLA-DR-positive microglial clusters over plaques were P2RY12-negative except for single cells observed within the cluster (arrows). ( G , H ). Interaction of P2RY12-immunoreactive microglia (purple) and Aβ plaques (brown). The panels show two types of interactions of P2RY12-positive microglia with plaques. Positive microglia are not present in close association with mature cored plaque ( G ), while they are present in close association with diffuse type of plaques ( H ). Specificity controls for P2RY12 staining of microglia. ( I , J ). Staining of representative sections with P2RY12 (Novus) antibody preabsorbed with immunizing peptide (I, +Pep) compared to staining of matched section with P2RY12 antibody non-absorbed ( J , -Pep). ( K , L ). Staining of matched sections with alternative P2RY12 antibody. Same staining pattern of microglia revealed with P2RY12 (Novus) antibody ( C ) as with P2RY12 (Alomone Labs) antibody (D ). Sections reacted with Alomone Lab P2RY12 required antigen retrieval to obtain positive staining pattern. All sections shown had been counterstained with neutral red to identify nuclei (red color). Abbreviations: ND: non-demented. AD: Alzheimer’s disease. MTG: middle temporal gyrus.—Pep: antibody without immunizing peptide. + Pep: antibody with immunizing peptide. Scale bars represent 50 μm.

    Journal: International Journal of Molecular Sciences

    Article Title: Patterns of Expression of Purinergic Receptor P2RY12, a Putative Marker for Non-Activated Microglia, in Aged and Alzheimer’s Disease Brains

    doi: 10.3390/ijms21020678

    Figure Lengend Snippet: Features of P2RY12-immunoreactive microglia. ( A , B ). Morphology of P2RY12-immunoreactive microglia (purple) in a low plaque non-demented (LPND) case ( A ) and AD case ( B ). Sections of middle temporal gyrus (MTG) were single-stained with antibody to P2RY12. Red arrows in panel B illustrate the lack of P2RY12 immunoreactive cells in an area occupied by plaque. ( C , D ). P2RY12-immunoreactive microglia (purple) are a feature in hippocampus sections from non-demented (ND) case ( H ) and Alzheimer’s disease (AD) case. Section shows staining in CA2 region of hippocampus. Continued presence of P2RY12-positive microglia in AD hippocampus ( D ) was noticeable. ( E , F ). Double-staining of section of ND and AD case with P2RY12 (purple) and HLA-DR (brown) showed limited overlap. HLA-DR-positive microglial clusters over plaques were P2RY12-negative except for single cells observed within the cluster (arrows). ( G , H ). Interaction of P2RY12-immunoreactive microglia (purple) and Aβ plaques (brown). The panels show two types of interactions of P2RY12-positive microglia with plaques. Positive microglia are not present in close association with mature cored plaque ( G ), while they are present in close association with diffuse type of plaques ( H ). Specificity controls for P2RY12 staining of microglia. ( I , J ). Staining of representative sections with P2RY12 (Novus) antibody preabsorbed with immunizing peptide (I, +Pep) compared to staining of matched section with P2RY12 antibody non-absorbed ( J , -Pep). ( K , L ). Staining of matched sections with alternative P2RY12 antibody. Same staining pattern of microglia revealed with P2RY12 (Novus) antibody ( C ) as with P2RY12 (Alomone Labs) antibody (D ). Sections reacted with Alomone Lab P2RY12 required antigen retrieval to obtain positive staining pattern. All sections shown had been counterstained with neutral red to identify nuclei (red color). Abbreviations: ND: non-demented. AD: Alzheimer’s disease. MTG: middle temporal gyrus.—Pep: antibody without immunizing peptide. + Pep: antibody with immunizing peptide. Scale bars represent 50 μm.

    Article Snippet: In addition, comparisons of immunostaining patterns were carried using an independent antibody to P2RY12 (1:250, Alomone Labs), prepared against an 18-amino acid peptide sequence that did not overlap with the immunizing sequence of the Novus antibody.

    Techniques: Staining, Double Staining

    Expression of P2RY12 mRNA and protein by in vitro cultures of human microglia. ( A ). Interleukin-4 stimulates P2RY12 mRNA expression. Bar chart showing results real time PCR analysis for P2RY12 mRNA of human microglia stimulated with indicated agents. Results of analysis of single human microglia case (each in triplicate) and representative of other analyses. Abbreviations: Con, control unstimulated: IL-4, interleukin-4 (40 ng/mL): Aβ2 and Aββ5 (Aβ (1–42) 2 μM and 5 μM): IFNγ, interferon-γ (100 ng/mL): LPS, lipopolysaccharide (100 ng/mL): LPS/IFNγ (doses combined): IL-6, interleukin-6 (40 ng/mL). **** p

    Journal: International Journal of Molecular Sciences

    Article Title: Patterns of Expression of Purinergic Receptor P2RY12, a Putative Marker for Non-Activated Microglia, in Aged and Alzheimer’s Disease Brains

    doi: 10.3390/ijms21020678

    Figure Lengend Snippet: Expression of P2RY12 mRNA and protein by in vitro cultures of human microglia. ( A ). Interleukin-4 stimulates P2RY12 mRNA expression. Bar chart showing results real time PCR analysis for P2RY12 mRNA of human microglia stimulated with indicated agents. Results of analysis of single human microglia case (each in triplicate) and representative of other analyses. Abbreviations: Con, control unstimulated: IL-4, interleukin-4 (40 ng/mL): Aβ2 and Aββ5 (Aβ (1–42) 2 μM and 5 μM): IFNγ, interferon-γ (100 ng/mL): LPS, lipopolysaccharide (100 ng/mL): LPS/IFNγ (doses combined): IL-6, interleukin-6 (40 ng/mL). **** p

    Article Snippet: In addition, comparisons of immunostaining patterns were carried using an independent antibody to P2RY12 (1:250, Alomone Labs), prepared against an 18-amino acid peptide sequence that did not overlap with the immunizing sequence of the Novus antibody.

    Techniques: Expressing, In Vitro, Real-time Polymerase Chain Reaction

    Confocal microscopy of P2RY12 and HLA-DR positive microglia in pathologically staged cases. ( A – I ) Images of P2RY12 (green), HLA-DR (red) and merged (yellow) with DAPI (blue) in MTG of LPND ( A – C ), HPND ( D – F ) and AD case ( G – I ) to show the distribution of P2RY12 and HLA-DR immunoreactivity. Examples of colocalization (yellow arrows) are shown in Merge image. Scale bar represents 50 μm.

    Journal: International Journal of Molecular Sciences

    Article Title: Patterns of Expression of Purinergic Receptor P2RY12, a Putative Marker for Non-Activated Microglia, in Aged and Alzheimer’s Disease Brains

    doi: 10.3390/ijms21020678

    Figure Lengend Snippet: Confocal microscopy of P2RY12 and HLA-DR positive microglia in pathologically staged cases. ( A – I ) Images of P2RY12 (green), HLA-DR (red) and merged (yellow) with DAPI (blue) in MTG of LPND ( A – C ), HPND ( D – F ) and AD case ( G – I ) to show the distribution of P2RY12 and HLA-DR immunoreactivity. Examples of colocalization (yellow arrows) are shown in Merge image. Scale bar represents 50 μm.

    Article Snippet: In addition, comparisons of immunostaining patterns were carried using an independent antibody to P2RY12 (1:250, Alomone Labs), prepared against an 18-amino acid peptide sequence that did not overlap with the immunizing sequence of the Novus antibody.

    Techniques: Confocal Microscopy

    Features of P2RY12-positive microglia interacting with phosphorylated tau-containing structures. ( A–F ). Dual-color DAB enzyme histochemistry illustrating different features of P2RY12-positive microglia (purple) with phosphorylated tau-positive structures (brown). Abbreviations: LPND; low plaque non-demented. HPND; high plaque non-demented. AD; Alzheimer’s disease. MTG; middle temporal gyrus. Scale bars represent 50 μm. ( G – I ). Dual-color laser confocal histochemistry showing interaction of P2RY12 positive microglia with early intracellular tangle in LPND case ( G ). Features of P2RY12-positive microglia interacting with mature tangle ( H ) and neuritic plaque ( I ) in AD cases. Scale bars represent 50 μm.

    Journal: International Journal of Molecular Sciences

    Article Title: Patterns of Expression of Purinergic Receptor P2RY12, a Putative Marker for Non-Activated Microglia, in Aged and Alzheimer’s Disease Brains

    doi: 10.3390/ijms21020678

    Figure Lengend Snippet: Features of P2RY12-positive microglia interacting with phosphorylated tau-containing structures. ( A–F ). Dual-color DAB enzyme histochemistry illustrating different features of P2RY12-positive microglia (purple) with phosphorylated tau-positive structures (brown). Abbreviations: LPND; low plaque non-demented. HPND; high plaque non-demented. AD; Alzheimer’s disease. MTG; middle temporal gyrus. Scale bars represent 50 μm. ( G – I ). Dual-color laser confocal histochemistry showing interaction of P2RY12 positive microglia with early intracellular tangle in LPND case ( G ). Features of P2RY12-positive microglia interacting with mature tangle ( H ) and neuritic plaque ( I ) in AD cases. Scale bars represent 50 μm.

    Article Snippet: In addition, comparisons of immunostaining patterns were carried using an independent antibody to P2RY12 (1:250, Alomone Labs), prepared against an 18-amino acid peptide sequence that did not overlap with the immunizing sequence of the Novus antibody.

    Techniques:

    Different microglial morphologies associated with P2RY12 expression. Representative immunohistochemistry results of tissue sections stained to identify P2RY12 (purple) alone and Aβ (brown), IBA-1 or phosphorylated tau. ( A ). Ramified microglia in LPND case. ( B , C ). Microglia with fragmented morphology in HPND cases. ( C ). Fragmented microglia associated with diffuse Aβ plaques. ( D ). P2RY12 microglia with tufted morphology in AD case. ( E , F ). Colocalization of P2RY12 and IBA-1. ( E ). Rod shaped IBA-1-positive microglia (brown arrow) with minimal P2RY12 immunoreactivity. P2RY12-positive, IBA-1-positive microglia (arrowheads ( E , F )). All P2RY12 immunoreactive microglia showed some IBA-1 immunoreactivity. ( F ). IBA-1 positive cluster surrounded by P2RY12 microglia. ( G – I ). Different morphologies of P2RY12-positive microglia interacting with Aβ plaques. ( G ). P2RY12-positive microglia with long processes interacting with diffuse plaques in HPND case. ( H , I ). P2RY12-positve microglia with activated morphologies (large cell bodies, short processes) interacting with dense Aβ plaques. ( J , K ). P2RY12-positive rod-shaped microglia in LPND ( J ) and HPND ( K ) sections. ( L ) P2RY12-positive microglial processes show interactions with neurons. Scale bars represent 50 μm.

    Journal: International Journal of Molecular Sciences

    Article Title: Patterns of Expression of Purinergic Receptor P2RY12, a Putative Marker for Non-Activated Microglia, in Aged and Alzheimer’s Disease Brains

    doi: 10.3390/ijms21020678

    Figure Lengend Snippet: Different microglial morphologies associated with P2RY12 expression. Representative immunohistochemistry results of tissue sections stained to identify P2RY12 (purple) alone and Aβ (brown), IBA-1 or phosphorylated tau. ( A ). Ramified microglia in LPND case. ( B , C ). Microglia with fragmented morphology in HPND cases. ( C ). Fragmented microglia associated with diffuse Aβ plaques. ( D ). P2RY12 microglia with tufted morphology in AD case. ( E , F ). Colocalization of P2RY12 and IBA-1. ( E ). Rod shaped IBA-1-positive microglia (brown arrow) with minimal P2RY12 immunoreactivity. P2RY12-positive, IBA-1-positive microglia (arrowheads ( E , F )). All P2RY12 immunoreactive microglia showed some IBA-1 immunoreactivity. ( F ). IBA-1 positive cluster surrounded by P2RY12 microglia. ( G – I ). Different morphologies of P2RY12-positive microglia interacting with Aβ plaques. ( G ). P2RY12-positive microglia with long processes interacting with diffuse plaques in HPND case. ( H , I ). P2RY12-positve microglia with activated morphologies (large cell bodies, short processes) interacting with dense Aβ plaques. ( J , K ). P2RY12-positive rod-shaped microglia in LPND ( J ) and HPND ( K ) sections. ( L ) P2RY12-positive microglial processes show interactions with neurons. Scale bars represent 50 μm.

    Article Snippet: In addition, comparisons of immunostaining patterns were carried using an independent antibody to P2RY12 (1:250, Alomone Labs), prepared against an 18-amino acid peptide sequence that did not overlap with the immunizing sequence of the Novus antibody.

    Techniques: Expressing, Immunohistochemistry, Staining

    Confocal microscopy of P2RY12 and CD68-positive microglia in pathologically staged cases. A – C). Low magnification merged images of P2RY12 (green), CD68 (red) and DAPI (blue) in MTG of LPND ( A ), HPND ( B ) and AD cases ( C ) to show the distribution of P2RY12 and CD68 immunoreactivity through cortical layer. Scale bars represent 50 μm. ( D – F ). Higher magnification merged images of CD68 (red) in MTG of LPND ( D ), HPND ( E ) and AD cases ( F ) to show the distribution of CD68 immunoreactivity. Similar amounts of CD68 immunoreactivity was present in each disease group. Scale bars represent 50 μm. ( G – I ). Merged images of P2RY12 (green) with the CD68 (red) images shown in ( D , E )) with DAPI (blue). Scale bars represent 50 μm.

    Journal: International Journal of Molecular Sciences

    Article Title: Patterns of Expression of Purinergic Receptor P2RY12, a Putative Marker for Non-Activated Microglia, in Aged and Alzheimer’s Disease Brains

    doi: 10.3390/ijms21020678

    Figure Lengend Snippet: Confocal microscopy of P2RY12 and CD68-positive microglia in pathologically staged cases. A – C). Low magnification merged images of P2RY12 (green), CD68 (red) and DAPI (blue) in MTG of LPND ( A ), HPND ( B ) and AD cases ( C ) to show the distribution of P2RY12 and CD68 immunoreactivity through cortical layer. Scale bars represent 50 μm. ( D – F ). Higher magnification merged images of CD68 (red) in MTG of LPND ( D ), HPND ( E ) and AD cases ( F ) to show the distribution of CD68 immunoreactivity. Similar amounts of CD68 immunoreactivity was present in each disease group. Scale bars represent 50 μm. ( G – I ). Merged images of P2RY12 (green) with the CD68 (red) images shown in ( D , E )) with DAPI (blue). Scale bars represent 50 μm.

    Article Snippet: In addition, comparisons of immunostaining patterns were carried using an independent antibody to P2RY12 (1:250, Alomone Labs), prepared against an 18-amino acid peptide sequence that did not overlap with the immunizing sequence of the Novus antibody.

    Techniques: Confocal Microscopy

    Quantitative polymerase chain reaction (qPCR) ΔCt analysis for hyperpolarization-activated cyclic nucleotide-gated channel (HCN channel) subunits in proprioceptive DRG cells. (A) Box and whisker plot show group data comparing ΔCt for GFP expression in GFP+ (green) and GFP− (blue) samples (normalized to β actin expression). The low GFP ΔCt in GFP+, but not GFP−, confirm high expression in this sample and the reliability of this sampling approach. (B) Box whisker plots plot shows group data comparing ΔCt values for HCN1–4 in GFP+ (green) and GFP− (blue) samples. The expression of all HCN subunits was significantly different between GFP+ and GFP− groups (where * p

    Journal: Frontiers in Molecular Neuroscience

    Article Title: Functional and Molecular Analysis of Proprioceptive Sensory Neuron Excitability in Mice

    doi: 10.3389/fnmol.2020.00036

    Figure Lengend Snippet: Quantitative polymerase chain reaction (qPCR) ΔCt analysis for hyperpolarization-activated cyclic nucleotide-gated channel (HCN channel) subunits in proprioceptive DRG cells. (A) Box and whisker plot show group data comparing ΔCt for GFP expression in GFP+ (green) and GFP− (blue) samples (normalized to β actin expression). The low GFP ΔCt in GFP+, but not GFP−, confirm high expression in this sample and the reliability of this sampling approach. (B) Box whisker plots plot shows group data comparing ΔCt values for HCN1–4 in GFP+ (green) and GFP− (blue) samples. The expression of all HCN subunits was significantly different between GFP+ and GFP− groups (where * p

    Article Snippet: The sections were then incubated in a cocktail of primary antibodies containing guinea pig anti-PV (1:500; Frontier Institute Cat# PV-GP-Af1000, RRID:AB_2336938 ) with either rabbit anti HCN1 (1:250; Alomone Labs Cat# APC-056, RRID:AB_2039900 ), rabbit anti-HCN2 (1:500; Alomone Labs Cat# APC-030, RRID:AB_2313726 ) or mouse anti-HCN4 (diluted 1:500; UC Davis/NIH NeuroMab Facility Cat# 73–150, RRID:AB_10673158).

    Techniques: Real-time Polymerase Chain Reaction, Whisker Assay, Expressing, Sampling

    Immunohistochemical localization of HCN subunits in PV-expressing DRG neurons. (A,B) PV-IR DRG neurons (green) in wild-type mice were shown to express both HCN1 ( A ; red) and HCN2 ( B ; red), with examples denoted by double arrowheads. (C,D) In DRGs from PVeGFP mice, GFP-expressing cells (green) showed immunolabeling for both HCN2 ( C ; red) and HCN4 ( D ; red). Immunolabeling for each of the HCN subunits was restricted to the cell membrane. Scale Bars (μm): (A,B) = 50; (C,D) = 10.

    Journal: Frontiers in Molecular Neuroscience

    Article Title: Functional and Molecular Analysis of Proprioceptive Sensory Neuron Excitability in Mice

    doi: 10.3389/fnmol.2020.00036

    Figure Lengend Snippet: Immunohistochemical localization of HCN subunits in PV-expressing DRG neurons. (A,B) PV-IR DRG neurons (green) in wild-type mice were shown to express both HCN1 ( A ; red) and HCN2 ( B ; red), with examples denoted by double arrowheads. (C,D) In DRGs from PVeGFP mice, GFP-expressing cells (green) showed immunolabeling for both HCN2 ( C ; red) and HCN4 ( D ; red). Immunolabeling for each of the HCN subunits was restricted to the cell membrane. Scale Bars (μm): (A,B) = 50; (C,D) = 10.

    Article Snippet: The sections were then incubated in a cocktail of primary antibodies containing guinea pig anti-PV (1:500; Frontier Institute Cat# PV-GP-Af1000, RRID:AB_2336938 ) with either rabbit anti HCN1 (1:250; Alomone Labs Cat# APC-056, RRID:AB_2039900 ), rabbit anti-HCN2 (1:500; Alomone Labs Cat# APC-030, RRID:AB_2313726 ) or mouse anti-HCN4 (diluted 1:500; UC Davis/NIH NeuroMab Facility Cat# 73–150, RRID:AB_10673158).

    Techniques: Immunohistochemistry, Expressing, Mouse Assay, Immunolabeling

    Inhibition of PKG prevented the proliferation of SVZ cells stimulated by PDE5 inhibitors. Cell proliferation following treatment with 1 μ M KT5823 and 1 μ M T0156 (a, d), 1 μ M sildenafil (b, e), or 10 μ M zaprinast (c, f) was assessed by incorporation of EdU and evaluated by flow cytometry, following 6 h or 24 h of treatment. Data are expressed as means ± SEM of at least 4 independent experiments. One-way ANOVA (Bonferroni's post-test), * P

    Journal: Stem Cells International

    Article Title: Stimulation of Neural Stem Cell Proliferation by Inhibition of Phosphodiesterase 5

    doi: 10.1155/2014/878397

    Figure Lengend Snippet: Inhibition of PKG prevented the proliferation of SVZ cells stimulated by PDE5 inhibitors. Cell proliferation following treatment with 1 μ M KT5823 and 1 μ M T0156 (a, d), 1 μ M sildenafil (b, e), or 10 μ M zaprinast (c, f) was assessed by incorporation of EdU and evaluated by flow cytometry, following 6 h or 24 h of treatment. Data are expressed as means ± SEM of at least 4 independent experiments. One-way ANOVA (Bonferroni's post-test), * P

    Article Snippet: KT5823 was obtained from Alomone Labs (Jerusalem, Israel) and 3-(5′-Hydroxymethyl-2′-furyl)-1-benzylindazole (YC-1) from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

    Techniques: Inhibition, Flow Cytometry, Cytometry

    Activation of sGC stimulates proliferation of NSC, increases ERK1/2 phosphorylation, and decreases p27 Kip1 levels. Cell proliferation following treatment with 20 μ M YC-1 (a) for 24 h was assessed by the incorporation of EdU and analyzed by flow cytometry. Levels of phospho-ERK1/2 following treatment with 20 μ M YC-1 (b) or YC-1 plus 1 μ M KT5823 (c) and p27 Kip1 levels following treatment with YC-1 (d) or YC-1 plus 1 μ M U0126 (e) or KT5823 (f) for 2 h were assessed by Western blot. Representative images are shown. Data are expressed as means ± SEM of at least 4 independent experiments. (a), (b), and (d) two-tailed t -test, * P

    Journal: Stem Cells International

    Article Title: Stimulation of Neural Stem Cell Proliferation by Inhibition of Phosphodiesterase 5

    doi: 10.1155/2014/878397

    Figure Lengend Snippet: Activation of sGC stimulates proliferation of NSC, increases ERK1/2 phosphorylation, and decreases p27 Kip1 levels. Cell proliferation following treatment with 20 μ M YC-1 (a) for 24 h was assessed by the incorporation of EdU and analyzed by flow cytometry. Levels of phospho-ERK1/2 following treatment with 20 μ M YC-1 (b) or YC-1 plus 1 μ M KT5823 (c) and p27 Kip1 levels following treatment with YC-1 (d) or YC-1 plus 1 μ M U0126 (e) or KT5823 (f) for 2 h were assessed by Western blot. Representative images are shown. Data are expressed as means ± SEM of at least 4 independent experiments. (a), (b), and (d) two-tailed t -test, * P

    Article Snippet: KT5823 was obtained from Alomone Labs (Jerusalem, Israel) and 3-(5′-Hydroxymethyl-2′-furyl)-1-benzylindazole (YC-1) from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

    Techniques: Activation Assay, Flow Cytometry, Cytometry, Western Blot, Two Tailed Test

    Inhibition of PKG did not prevent the decrease of p27 Kip1 levels by T0156. p27 Kip1 levels following treatment with 1 μ M KT5823 and 1 μ M T0156 (a), 1 μ M sildenafil (b), or 10 μ M zaprinast (c) for 2 h were assessed by Western blot. Representative images are shown. Data are expressed as means ± SEM of at least 4 independent experiments. One-way ANOVA (Bonferroni's post-test), * P

    Journal: Stem Cells International

    Article Title: Stimulation of Neural Stem Cell Proliferation by Inhibition of Phosphodiesterase 5

    doi: 10.1155/2014/878397

    Figure Lengend Snippet: Inhibition of PKG did not prevent the decrease of p27 Kip1 levels by T0156. p27 Kip1 levels following treatment with 1 μ M KT5823 and 1 μ M T0156 (a), 1 μ M sildenafil (b), or 10 μ M zaprinast (c) for 2 h were assessed by Western blot. Representative images are shown. Data are expressed as means ± SEM of at least 4 independent experiments. One-way ANOVA (Bonferroni's post-test), * P

    Article Snippet: KT5823 was obtained from Alomone Labs (Jerusalem, Israel) and 3-(5′-Hydroxymethyl-2′-furyl)-1-benzylindazole (YC-1) from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

    Techniques: Inhibition, Western Blot

    Inhibition of PKG prevented the phosphorylation of ERK1/2 by treatment with T0156 or zaprinast. Levels of phospho-ERK1/2 following treatment with 1 μ M KT5823 and 1 μ M T0156 (a), 1 μ M sildenafil (b), and 10 μ M zaprinast (c) for 2 h were assessed by Western blot. Representative images are shown. Data are expressed as means ± SEM of at least 4 independent experiments. One-way ANOVA (Bonferroni's post-test), * P

    Journal: Stem Cells International

    Article Title: Stimulation of Neural Stem Cell Proliferation by Inhibition of Phosphodiesterase 5

    doi: 10.1155/2014/878397

    Figure Lengend Snippet: Inhibition of PKG prevented the phosphorylation of ERK1/2 by treatment with T0156 or zaprinast. Levels of phospho-ERK1/2 following treatment with 1 μ M KT5823 and 1 μ M T0156 (a), 1 μ M sildenafil (b), and 10 μ M zaprinast (c) for 2 h were assessed by Western blot. Representative images are shown. Data are expressed as means ± SEM of at least 4 independent experiments. One-way ANOVA (Bonferroni's post-test), * P

    Article Snippet: KT5823 was obtained from Alomone Labs (Jerusalem, Israel) and 3-(5′-Hydroxymethyl-2′-furyl)-1-benzylindazole (YC-1) from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

    Techniques: Inhibition, Western Blot

    Line-blot for qualitative analysis of anti-BDNF antibodies specificity. ( A ) The antibodies from each ELISA kit were tested for specificity against pro-BDNF or mature BDNF. The BDNF standards blotted were commercial pro-BDNF (Alomone; 10 pg/lane), mature BDNF (1 and 2 from Alomone and Sigma, respectively; both 1000 pg/lane) and the standard BDNF protein included in each kit (Aviscera-Bioscience and Biosensis: 10 pg/lane; Millipore-ChemiKine TM , Millipore-Milliplex ® - and R D System-Quantikine ® : 100 pg/lane; Promega-Emax ® : 1000 pg/lane). BSA (1000 pg/lane) was used as a negative control. The mouse monoclonal anti-BDNF antibody, (1:1000; Sigma) was tested as a control. ( B ) Central region of the same blot shown in A, from an overexposed film to better visualize the reactivity against pro-BDNF. ( C ) Reactivity of antibodies from Biosensis, Promega-Emax ® pAb and Sigma on a dot blot in which the same quantity of pro-BNDF and mature BDNF were spotted (100 pg each). Each antibody from the ELISA kits was used at the dilution suggested by the manufacturer’s instructions. mAb: Promega-Emax ® monoclonal capture antibody for plate coating. pAb: Promega-Emax ® polyclonal detection antibody.

    Journal: Scientific Reports

    Article Title: A method for reproducible measurements of serum BDNF: comparison of the performance of six commercial assays

    doi: 10.1038/srep17989

    Figure Lengend Snippet: Line-blot for qualitative analysis of anti-BDNF antibodies specificity. ( A ) The antibodies from each ELISA kit were tested for specificity against pro-BDNF or mature BDNF. The BDNF standards blotted were commercial pro-BDNF (Alomone; 10 pg/lane), mature BDNF (1 and 2 from Alomone and Sigma, respectively; both 1000 pg/lane) and the standard BDNF protein included in each kit (Aviscera-Bioscience and Biosensis: 10 pg/lane; Millipore-ChemiKine TM , Millipore-Milliplex ® - and R D System-Quantikine ® : 100 pg/lane; Promega-Emax ® : 1000 pg/lane). BSA (1000 pg/lane) was used as a negative control. The mouse monoclonal anti-BDNF antibody, (1:1000; Sigma) was tested as a control. ( B ) Central region of the same blot shown in A, from an overexposed film to better visualize the reactivity against pro-BDNF. ( C ) Reactivity of antibodies from Biosensis, Promega-Emax ® pAb and Sigma on a dot blot in which the same quantity of pro-BNDF and mature BDNF were spotted (100 pg each). Each antibody from the ELISA kits was used at the dilution suggested by the manufacturer’s instructions. mAb: Promega-Emax ® monoclonal capture antibody for plate coating. pAb: Promega-Emax ® polyclonal detection antibody.

    Article Snippet: Of note, Aviscera-Bioscience and R & D System-Quantikine® kits were claimed to be specific for mature BDNF and to have some cross-reactivity with the human pro-BDNF.

    Techniques: Enzyme-linked Immunosorbent Assay, Negative Control, Dot Blot