rabbit anti human monoclonal antibodies against eta  (Alomone Labs)


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    Alomone Labs rabbit anti human monoclonal antibodies against eta
    Immunostaining of sections of stenotic valve from a patient with rheumatic fever. Top left. ET-1 staining associated with fibrotic regions and neovascularization. Top middle. <t>ETA</t> receptor staining of fibrotic region and neovascularization. Top right. H + E-stained section. Lower left. No ET-3 immunostaining. Lower <t>middle.</t> <t>ETB</t> receptor immunostaining of fibrotic region and neovascularization. Lower right. Negative control. ET=endothelin; ETA=endothelin receptor A; ETB=endothelin receptor B
    Rabbit Anti Human Monoclonal Antibodies Against Eta, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti human monoclonal antibodies against eta/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti human monoclonal antibodies against eta - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "Analysis of immunostaining and western blotting of endothelin 1 and its receptors in mitral stenosis"

    Article Title: Analysis of immunostaining and western blotting of endothelin 1 and its receptors in mitral stenosis

    Journal: Revista Brasileira de Cirurgia Cardiovascular : órgão oficial da Sociedade Brasileira de Cirurgia Cardiovascular

    doi: 10.5935/1678-9741.20150004

    Immunostaining of sections of stenotic valve from a patient with rheumatic fever. Top left. ET-1 staining associated with fibrotic regions and neovascularization. Top middle. ETA receptor staining of fibrotic region and neovascularization. Top right. H + E-stained section. Lower left. No ET-3 immunostaining. Lower middle. ETB receptor immunostaining of fibrotic region and neovascularization. Lower right. Negative control. ET=endothelin; ETA=endothelin receptor A; ETB=endothelin receptor B
    Figure Legend Snippet: Immunostaining of sections of stenotic valve from a patient with rheumatic fever. Top left. ET-1 staining associated with fibrotic regions and neovascularization. Top middle. ETA receptor staining of fibrotic region and neovascularization. Top right. H + E-stained section. Lower left. No ET-3 immunostaining. Lower middle. ETB receptor immunostaining of fibrotic region and neovascularization. Lower right. Negative control. ET=endothelin; ETA=endothelin receptor A; ETB=endothelin receptor B

    Techniques Used: Immunostaining, Staining, Negative Control

    rabbit anti human monoclonal antibodies against eta  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
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  • 94

    Structured Review

    Alomone Labs rabbit anti human monoclonal antibodies against eta
    Immunostaining of sections of stenotic valve from a patient with rheumatic fever. Top left. ET-1 staining associated with fibrotic regions and neovascularization. Top middle. <t>ETA</t> receptor staining of fibrotic region and neovascularization. Top right. H + E-stained section. Lower left. No ET-3 immunostaining. Lower <t>middle.</t> <t>ETB</t> receptor immunostaining of fibrotic region and neovascularization. Lower right. Negative control. ET=endothelin; ETA=endothelin receptor A; ETB=endothelin receptor B
    Rabbit Anti Human Monoclonal Antibodies Against Eta, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti human monoclonal antibodies against eta/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti human monoclonal antibodies against eta - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "Analysis of immunostaining and western blotting of endothelin 1 and its receptors in mitral stenosis"

    Article Title: Analysis of immunostaining and western blotting of endothelin 1 and its receptors in mitral stenosis

    Journal: Revista Brasileira de Cirurgia Cardiovascular : órgão oficial da Sociedade Brasileira de Cirurgia Cardiovascular

    doi: 10.5935/1678-9741.20150004

    Immunostaining of sections of stenotic valve from a patient with rheumatic fever. Top left. ET-1 staining associated with fibrotic regions and neovascularization. Top middle. ETA receptor staining of fibrotic region and neovascularization. Top right. H + E-stained section. Lower left. No ET-3 immunostaining. Lower middle. ETB receptor immunostaining of fibrotic region and neovascularization. Lower right. Negative control. ET=endothelin; ETA=endothelin receptor A; ETB=endothelin receptor B
    Figure Legend Snippet: Immunostaining of sections of stenotic valve from a patient with rheumatic fever. Top left. ET-1 staining associated with fibrotic regions and neovascularization. Top middle. ETA receptor staining of fibrotic region and neovascularization. Top right. H + E-stained section. Lower left. No ET-3 immunostaining. Lower middle. ETB receptor immunostaining of fibrotic region and neovascularization. Lower right. Negative control. ET=endothelin; ETA=endothelin receptor A; ETB=endothelin receptor B

    Techniques Used: Immunostaining, Staining, Negative Control

    rabbit igg anti p2y12r  (Alomone Labs)


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    Alomone Labs rabbit igg anti p2y12r
    A Immunohistostainings of post-mortem human hemispheric white matter tissue (healthy donor) showing microglial cells (blue, resident microglia, TMEM119) contacting nodes of Ranvier (Na V , brown). B , C Immunofluorescent stainings of post-mortem human hemispheric white matter tissue (healthy donor) showing microglial cells (green; B , resident microglia, TMEM119, and C , homeostatic microglia, <t>P2Y12R)</t> contacting nodes of Ranvier (Na V , red). B ii, C ii 3D reconstructions correspond to the boxed area in B i and C i respectively. Scale bars: (2D) A , B , C 10 μm; (3D), B , C 2 μm. A : n = 6 samples, B , C : n = 5 samples.
    Rabbit Igg Anti P2y12r, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit igg anti p2y12r/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit igg anti p2y12r - by Bioz Stars, 2023-01
    93/100 stars

    Images

    1) Product Images from "Microglia-neuron interaction at nodes of Ranvier depends on neuronal activity through potassium release and contributes to remyelination"

    Article Title: Microglia-neuron interaction at nodes of Ranvier depends on neuronal activity through potassium release and contributes to remyelination

    Journal: Nature Communications

    doi: 10.1038/s41467-021-25486-7

    A Immunohistostainings of post-mortem human hemispheric white matter tissue (healthy donor) showing microglial cells (blue, resident microglia, TMEM119) contacting nodes of Ranvier (Na V , brown). B , C Immunofluorescent stainings of post-mortem human hemispheric white matter tissue (healthy donor) showing microglial cells (green; B , resident microglia, TMEM119, and C , homeostatic microglia, P2Y12R) contacting nodes of Ranvier (Na V , red). B ii, C ii 3D reconstructions correspond to the boxed area in B i and C i respectively. Scale bars: (2D) A , B , C 10 μm; (3D), B , C 2 μm. A : n = 6 samples, B , C : n = 5 samples.
    Figure Legend Snippet: A Immunohistostainings of post-mortem human hemispheric white matter tissue (healthy donor) showing microglial cells (blue, resident microglia, TMEM119) contacting nodes of Ranvier (Na V , brown). B , C Immunofluorescent stainings of post-mortem human hemispheric white matter tissue (healthy donor) showing microglial cells (green; B , resident microglia, TMEM119, and C , homeostatic microglia, P2Y12R) contacting nodes of Ranvier (Na V , red). B ii, C ii 3D reconstructions correspond to the boxed area in B i and C i respectively. Scale bars: (2D) A , B , C 10 μm; (3D), B , C 2 μm. A : n = 6 samples, B , C : n = 5 samples.

    Techniques Used:

    anti human anti adenosine a 2a r rabbit polyclonal antibody  (Alomone Labs)


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    Alomone Labs anti human anti adenosine a 2a r rabbit polyclonal antibody
    ADORA2A siRNAs decrease ADORA2A mRNA and A <t>2A</t> <t>R</t> protein expression (A) Representation of the structure of the lipid NPs labeled with fluorescent streptavidin and biotinylated targeting antibody and loaded with siRNAs. PE: phosphoethanolamine; PEG: polyethylene glycol (B) Relative quantity (RQ) of ADORA2A mRNA expression in activated HD CD3 + or CD8 + T cells 24 h post-transfection with 10 nM scr or ADORA2A siRNAs (n = 5; 3 CD3 + T cell samples, 2 CD8 + T cell samples). GAPDH (Glyceraldehyde 3-phosphate dehydrogenase) was used as the reference gene. Data are shown as mean ± standard deviation and significance is determined by a paired Student’s t test. (C) (Left) Fold change of normalized A 2A R protein expression in activated HD CD3 + or CD8 + T cells 24 h (n = 5; 3 CD3 + T cell samples, 2 CD8 + T cell samples), 48 h (n = 5; 3 CD3 + T cell samples, 2 CD8 + T cell samples), or 72 h (n = 4; 2 CD3 + T cell samples, 2 CD8 + T cell samples) post-transfection with 10 nM ADORA2A siRNAs as compared to scr-RNAs. (Right) The normalized MFI of A 2A R expression showing the highest degree of A 2A R protein knockdown post-transfection with 10 nM ADORA2A siRNAs as compared to corresponding scr-RNAs for each individual sample regardless of time point at which knockdown occurred. Data are shown as mean ± standard error of the mean (n = 5). Data are compared using a paired Student’s t test. (D) Flow cytometry gating strategy for determining A 2A R expression in transfected cells. The cells were first gated on T lymphocytes and then on GFP fluorescence (indicating that the cells were successfully transfected with siRNAs) to determine A 2A R expression. The unstained sample is indicated in orange in the histogram while the A 2A R expression of cells treated with scr-RNAs is indicated in blue and the A 2A R expression of cells treated with ADORA2A siRNAs is indicated in red. (E) Representative histogram showing A 2A R antibody specificity where CD3 + T cells were stained with A 2A R antibody ± peptide followed by secondary antibody. Unstained cells are indicated in gray, A 2A R antibody + peptide is indicated in red and A 2A R antibody alone is indicated in blue. Shown here is a representative experiment from two identical experiments performed in HDs.
    Anti Human Anti Adenosine A 2a R Rabbit Polyclonal Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti human anti adenosine a 2a r rabbit polyclonal antibody/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti human anti adenosine a 2a r rabbit polyclonal antibody - by Bioz Stars, 2023-01
    93/100 stars

    Images

    1) Product Images from "Targeted knockdown of the adenosine A 2A receptor by lipid NPs rescues the chemotaxis of head and neck cancer memory T cells"

    Article Title: Targeted knockdown of the adenosine A 2A receptor by lipid NPs rescues the chemotaxis of head and neck cancer memory T cells

    Journal: Molecular Therapy. Methods & Clinical Development

    doi: 10.1016/j.omtm.2021.03.001

    ADORA2A siRNAs decrease ADORA2A mRNA and A 2A R protein expression (A) Representation of the structure of the lipid NPs labeled with fluorescent streptavidin and biotinylated targeting antibody and loaded with siRNAs. PE: phosphoethanolamine; PEG: polyethylene glycol (B) Relative quantity (RQ) of ADORA2A mRNA expression in activated HD CD3 + or CD8 + T cells 24 h post-transfection with 10 nM scr or ADORA2A siRNAs (n = 5; 3 CD3 + T cell samples, 2 CD8 + T cell samples). GAPDH (Glyceraldehyde 3-phosphate dehydrogenase) was used as the reference gene. Data are shown as mean ± standard deviation and significance is determined by a paired Student’s t test. (C) (Left) Fold change of normalized A 2A R protein expression in activated HD CD3 + or CD8 + T cells 24 h (n = 5; 3 CD3 + T cell samples, 2 CD8 + T cell samples), 48 h (n = 5; 3 CD3 + T cell samples, 2 CD8 + T cell samples), or 72 h (n = 4; 2 CD3 + T cell samples, 2 CD8 + T cell samples) post-transfection with 10 nM ADORA2A siRNAs as compared to scr-RNAs. (Right) The normalized MFI of A 2A R expression showing the highest degree of A 2A R protein knockdown post-transfection with 10 nM ADORA2A siRNAs as compared to corresponding scr-RNAs for each individual sample regardless of time point at which knockdown occurred. Data are shown as mean ± standard error of the mean (n = 5). Data are compared using a paired Student’s t test. (D) Flow cytometry gating strategy for determining A 2A R expression in transfected cells. The cells were first gated on T lymphocytes and then on GFP fluorescence (indicating that the cells were successfully transfected with siRNAs) to determine A 2A R expression. The unstained sample is indicated in orange in the histogram while the A 2A R expression of cells treated with scr-RNAs is indicated in blue and the A 2A R expression of cells treated with ADORA2A siRNAs is indicated in red. (E) Representative histogram showing A 2A R antibody specificity where CD3 + T cells were stained with A 2A R antibody ± peptide followed by secondary antibody. Unstained cells are indicated in gray, A 2A R antibody + peptide is indicated in red and A 2A R antibody alone is indicated in blue. Shown here is a representative experiment from two identical experiments performed in HDs.
    Figure Legend Snippet: ADORA2A siRNAs decrease ADORA2A mRNA and A 2A R protein expression (A) Representation of the structure of the lipid NPs labeled with fluorescent streptavidin and biotinylated targeting antibody and loaded with siRNAs. PE: phosphoethanolamine; PEG: polyethylene glycol (B) Relative quantity (RQ) of ADORA2A mRNA expression in activated HD CD3 + or CD8 + T cells 24 h post-transfection with 10 nM scr or ADORA2A siRNAs (n = 5; 3 CD3 + T cell samples, 2 CD8 + T cell samples). GAPDH (Glyceraldehyde 3-phosphate dehydrogenase) was used as the reference gene. Data are shown as mean ± standard deviation and significance is determined by a paired Student’s t test. (C) (Left) Fold change of normalized A 2A R protein expression in activated HD CD3 + or CD8 + T cells 24 h (n = 5; 3 CD3 + T cell samples, 2 CD8 + T cell samples), 48 h (n = 5; 3 CD3 + T cell samples, 2 CD8 + T cell samples), or 72 h (n = 4; 2 CD3 + T cell samples, 2 CD8 + T cell samples) post-transfection with 10 nM ADORA2A siRNAs as compared to scr-RNAs. (Right) The normalized MFI of A 2A R expression showing the highest degree of A 2A R protein knockdown post-transfection with 10 nM ADORA2A siRNAs as compared to corresponding scr-RNAs for each individual sample regardless of time point at which knockdown occurred. Data are shown as mean ± standard error of the mean (n = 5). Data are compared using a paired Student’s t test. (D) Flow cytometry gating strategy for determining A 2A R expression in transfected cells. The cells were first gated on T lymphocytes and then on GFP fluorescence (indicating that the cells were successfully transfected with siRNAs) to determine A 2A R expression. The unstained sample is indicated in orange in the histogram while the A 2A R expression of cells treated with scr-RNAs is indicated in blue and the A 2A R expression of cells treated with ADORA2A siRNAs is indicated in red. (E) Representative histogram showing A 2A R antibody specificity where CD3 + T cells were stained with A 2A R antibody ± peptide followed by secondary antibody. Unstained cells are indicated in gray, A 2A R antibody + peptide is indicated in red and A 2A R antibody alone is indicated in blue. Shown here is a representative experiment from two identical experiments performed in HDs.

    Techniques Used: Expressing, Labeling, Transfection, Standard Deviation, Flow Cytometry, Fluorescence, Staining

    CD45RO(A 2A R)-NPs rescue the chemotactic ability of HNSCC CD8 + memory T cells in the presence of adenosine (A and B) Representative two point trajectories of T cells migrating along the CXCL10 gradient (green) or CXCL10 + adenosine (ADO) gradient (blue) in HNSCC CD8 + memory T cells treated with (A) CD45RO(scr)-NPs or (B) CD45RO(A 2A R)-NPs. Trajectories artificially set to start at the origin with the red triangle representing the Y-COM. (C) Y-COM of HNSCC CD8 + memory T cells treated with (left) CD45RO(scr)-NPs (n = 6) or (right) CD45RO(A 2A R)-NPs (n = 6) in the presence of CXCL10 or CXCL10 + adenosine. Significance was determined using (left) a Wilcoxon signed rank test or (right) paired Student’s t test. (D) Percent inhibition of Y-COM of HNSCC CD8 + memory T cells in the presence of CXCL10 + adenosine compared to CXCL10 alone when treated with CD45RO(scr)-NPs (n = 6) compared to CD45RO(A 2A R)-NPs (n = 6). Data are represented single dot plots (colored dots; wherein each dot represents a single donor) and as mean ± standard deviation (shown in black). Significance was determined using Student’s t test.
    Figure Legend Snippet: CD45RO(A 2A R)-NPs rescue the chemotactic ability of HNSCC CD8 + memory T cells in the presence of adenosine (A and B) Representative two point trajectories of T cells migrating along the CXCL10 gradient (green) or CXCL10 + adenosine (ADO) gradient (blue) in HNSCC CD8 + memory T cells treated with (A) CD45RO(scr)-NPs or (B) CD45RO(A 2A R)-NPs. Trajectories artificially set to start at the origin with the red triangle representing the Y-COM. (C) Y-COM of HNSCC CD8 + memory T cells treated with (left) CD45RO(scr)-NPs (n = 6) or (right) CD45RO(A 2A R)-NPs (n = 6) in the presence of CXCL10 or CXCL10 + adenosine. Significance was determined using (left) a Wilcoxon signed rank test or (right) paired Student’s t test. (D) Percent inhibition of Y-COM of HNSCC CD8 + memory T cells in the presence of CXCL10 + adenosine compared to CXCL10 alone when treated with CD45RO(scr)-NPs (n = 6) compared to CD45RO(A 2A R)-NPs (n = 6). Data are represented single dot plots (colored dots; wherein each dot represents a single donor) and as mean ± standard deviation (shown in black). Significance was determined using Student’s t test.

    Techniques Used: Inhibition, Standard Deviation

    anti human anti adenosine a 2a r rabbit polyclonal antibody  (Alomone Labs)


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    Alomone Labs anti human anti adenosine a 2a r rabbit polyclonal antibody
    ADORA2A siRNAs decrease ADORA2A mRNA and A <t>2A</t> <t>R</t> protein expression (A) Representation of the structure of the lipid NPs labeled with fluorescent streptavidin and biotinylated targeting antibody and loaded with siRNAs. PE: phosphoethanolamine; PEG: polyethylene glycol (B) Relative quantity (RQ) of ADORA2A mRNA expression in activated HD CD3 + or CD8 + T cells 24 h post-transfection with 10 nM scr or ADORA2A siRNAs (n = 5; 3 CD3 + T cell samples, 2 CD8 + T cell samples). GAPDH (Glyceraldehyde 3-phosphate dehydrogenase) was used as the reference gene. Data are shown as mean ± standard deviation and significance is determined by a paired Student’s t test. (C) (Left) Fold change of normalized A 2A R protein expression in activated HD CD3 + or CD8 + T cells 24 h (n = 5; 3 CD3 + T cell samples, 2 CD8 + T cell samples), 48 h (n = 5; 3 CD3 + T cell samples, 2 CD8 + T cell samples), or 72 h (n = 4; 2 CD3 + T cell samples, 2 CD8 + T cell samples) post-transfection with 10 nM ADORA2A siRNAs as compared to scr-RNAs. (Right) The normalized MFI of A 2A R expression showing the highest degree of A 2A R protein knockdown post-transfection with 10 nM ADORA2A siRNAs as compared to corresponding scr-RNAs for each individual sample regardless of time point at which knockdown occurred. Data are shown as mean ± standard error of the mean (n = 5). Data are compared using a paired Student’s t test. (D) Flow cytometry gating strategy for determining A 2A R expression in transfected cells. The cells were first gated on T lymphocytes and then on GFP fluorescence (indicating that the cells were successfully transfected with siRNAs) to determine A 2A R expression. The unstained sample is indicated in orange in the histogram while the A 2A R expression of cells treated with scr-RNAs is indicated in blue and the A 2A R expression of cells treated with ADORA2A siRNAs is indicated in red. (E) Representative histogram showing A 2A R antibody specificity where CD3 + T cells were stained with A 2A R antibody ± peptide followed by secondary antibody. Unstained cells are indicated in gray, A 2A R antibody + peptide is indicated in red and A 2A R antibody alone is indicated in blue. Shown here is a representative experiment from two identical experiments performed in HDs.
    Anti Human Anti Adenosine A 2a R Rabbit Polyclonal Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti human anti adenosine a 2a r rabbit polyclonal antibody/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti human anti adenosine a 2a r rabbit polyclonal antibody - by Bioz Stars, 2023-01
    93/100 stars

    Images

    1) Product Images from "Targeted knockdown of the adenosine A 2A receptor by lipid NPs rescues the chemotaxis of head and neck cancer memory T cells"

    Article Title: Targeted knockdown of the adenosine A 2A receptor by lipid NPs rescues the chemotaxis of head and neck cancer memory T cells

    Journal: Molecular Therapy. Methods & Clinical Development

    doi: 10.1016/j.omtm.2021.03.001

    ADORA2A siRNAs decrease ADORA2A mRNA and A 2A R protein expression (A) Representation of the structure of the lipid NPs labeled with fluorescent streptavidin and biotinylated targeting antibody and loaded with siRNAs. PE: phosphoethanolamine; PEG: polyethylene glycol (B) Relative quantity (RQ) of ADORA2A mRNA expression in activated HD CD3 + or CD8 + T cells 24 h post-transfection with 10 nM scr or ADORA2A siRNAs (n = 5; 3 CD3 + T cell samples, 2 CD8 + T cell samples). GAPDH (Glyceraldehyde 3-phosphate dehydrogenase) was used as the reference gene. Data are shown as mean ± standard deviation and significance is determined by a paired Student’s t test. (C) (Left) Fold change of normalized A 2A R protein expression in activated HD CD3 + or CD8 + T cells 24 h (n = 5; 3 CD3 + T cell samples, 2 CD8 + T cell samples), 48 h (n = 5; 3 CD3 + T cell samples, 2 CD8 + T cell samples), or 72 h (n = 4; 2 CD3 + T cell samples, 2 CD8 + T cell samples) post-transfection with 10 nM ADORA2A siRNAs as compared to scr-RNAs. (Right) The normalized MFI of A 2A R expression showing the highest degree of A 2A R protein knockdown post-transfection with 10 nM ADORA2A siRNAs as compared to corresponding scr-RNAs for each individual sample regardless of time point at which knockdown occurred. Data are shown as mean ± standard error of the mean (n = 5). Data are compared using a paired Student’s t test. (D) Flow cytometry gating strategy for determining A 2A R expression in transfected cells. The cells were first gated on T lymphocytes and then on GFP fluorescence (indicating that the cells were successfully transfected with siRNAs) to determine A 2A R expression. The unstained sample is indicated in orange in the histogram while the A 2A R expression of cells treated with scr-RNAs is indicated in blue and the A 2A R expression of cells treated with ADORA2A siRNAs is indicated in red. (E) Representative histogram showing A 2A R antibody specificity where CD3 + T cells were stained with A 2A R antibody ± peptide followed by secondary antibody. Unstained cells are indicated in gray, A 2A R antibody + peptide is indicated in red and A 2A R antibody alone is indicated in blue. Shown here is a representative experiment from two identical experiments performed in HDs.
    Figure Legend Snippet: ADORA2A siRNAs decrease ADORA2A mRNA and A 2A R protein expression (A) Representation of the structure of the lipid NPs labeled with fluorescent streptavidin and biotinylated targeting antibody and loaded with siRNAs. PE: phosphoethanolamine; PEG: polyethylene glycol (B) Relative quantity (RQ) of ADORA2A mRNA expression in activated HD CD3 + or CD8 + T cells 24 h post-transfection with 10 nM scr or ADORA2A siRNAs (n = 5; 3 CD3 + T cell samples, 2 CD8 + T cell samples). GAPDH (Glyceraldehyde 3-phosphate dehydrogenase) was used as the reference gene. Data are shown as mean ± standard deviation and significance is determined by a paired Student’s t test. (C) (Left) Fold change of normalized A 2A R protein expression in activated HD CD3 + or CD8 + T cells 24 h (n = 5; 3 CD3 + T cell samples, 2 CD8 + T cell samples), 48 h (n = 5; 3 CD3 + T cell samples, 2 CD8 + T cell samples), or 72 h (n = 4; 2 CD3 + T cell samples, 2 CD8 + T cell samples) post-transfection with 10 nM ADORA2A siRNAs as compared to scr-RNAs. (Right) The normalized MFI of A 2A R expression showing the highest degree of A 2A R protein knockdown post-transfection with 10 nM ADORA2A siRNAs as compared to corresponding scr-RNAs for each individual sample regardless of time point at which knockdown occurred. Data are shown as mean ± standard error of the mean (n = 5). Data are compared using a paired Student’s t test. (D) Flow cytometry gating strategy for determining A 2A R expression in transfected cells. The cells were first gated on T lymphocytes and then on GFP fluorescence (indicating that the cells were successfully transfected with siRNAs) to determine A 2A R expression. The unstained sample is indicated in orange in the histogram while the A 2A R expression of cells treated with scr-RNAs is indicated in blue and the A 2A R expression of cells treated with ADORA2A siRNAs is indicated in red. (E) Representative histogram showing A 2A R antibody specificity where CD3 + T cells were stained with A 2A R antibody ± peptide followed by secondary antibody. Unstained cells are indicated in gray, A 2A R antibody + peptide is indicated in red and A 2A R antibody alone is indicated in blue. Shown here is a representative experiment from two identical experiments performed in HDs.

    Techniques Used: Expressing, Labeling, Transfection, Standard Deviation, Flow Cytometry, Fluorescence, Staining

    CD45RO(A 2A R)-NPs rescue the chemotactic ability of HNSCC CD8 + memory T cells in the presence of adenosine (A and B) Representative two point trajectories of T cells migrating along the CXCL10 gradient (green) or CXCL10 + adenosine (ADO) gradient (blue) in HNSCC CD8 + memory T cells treated with (A) CD45RO(scr)-NPs or (B) CD45RO(A 2A R)-NPs. Trajectories artificially set to start at the origin with the red triangle representing the Y-COM. (C) Y-COM of HNSCC CD8 + memory T cells treated with (left) CD45RO(scr)-NPs (n = 6) or (right) CD45RO(A 2A R)-NPs (n = 6) in the presence of CXCL10 or CXCL10 + adenosine. Significance was determined using (left) a Wilcoxon signed rank test or (right) paired Student’s t test. (D) Percent inhibition of Y-COM of HNSCC CD8 + memory T cells in the presence of CXCL10 + adenosine compared to CXCL10 alone when treated with CD45RO(scr)-NPs (n = 6) compared to CD45RO(A 2A R)-NPs (n = 6). Data are represented single dot plots (colored dots; wherein each dot represents a single donor) and as mean ± standard deviation (shown in black). Significance was determined using Student’s t test.
    Figure Legend Snippet: CD45RO(A 2A R)-NPs rescue the chemotactic ability of HNSCC CD8 + memory T cells in the presence of adenosine (A and B) Representative two point trajectories of T cells migrating along the CXCL10 gradient (green) or CXCL10 + adenosine (ADO) gradient (blue) in HNSCC CD8 + memory T cells treated with (A) CD45RO(scr)-NPs or (B) CD45RO(A 2A R)-NPs. Trajectories artificially set to start at the origin with the red triangle representing the Y-COM. (C) Y-COM of HNSCC CD8 + memory T cells treated with (left) CD45RO(scr)-NPs (n = 6) or (right) CD45RO(A 2A R)-NPs (n = 6) in the presence of CXCL10 or CXCL10 + adenosine. Significance was determined using (left) a Wilcoxon signed rank test or (right) paired Student’s t test. (D) Percent inhibition of Y-COM of HNSCC CD8 + memory T cells in the presence of CXCL10 + adenosine compared to CXCL10 alone when treated with CD45RO(scr)-NPs (n = 6) compared to CD45RO(A 2A R)-NPs (n = 6). Data are represented single dot plots (colored dots; wherein each dot represents a single donor) and as mean ± standard deviation (shown in black). Significance was determined using Student’s t test.

    Techniques Used: Inhibition, Standard Deviation

    rabbit anti a2a  (Alomone Labs)


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    Alomone Labs rabbit anti a2a
    Interaction between <t>A2A</t> and D2 on rat striatal astrocyte processes: co-immunoprecipitation. ( A ) Aliquots (300 µg) of Triton X-100-soluble proteins obtained from gliosomes were immunoprecipitated with 1 µg of anti-A2A antibody as described in Methods. Immunoprecipitated (IP) and not immunoprecipitated (Output) materials were analyzed by immunoblotting using the anti-A2A antibody. IP and Output were also analyzed using anti-D2 antibody. A representative blot (of three) is shown. D2 immunoreactive bands were quantified and the data were reported in the graph. Values are means ± SEM ( n = 3). ( B ) Aliquots (300 µg) of Triton X-100-soluble proteins obtained from gliosomes were immunoprecipitated with 1 µg of anti-D2 antibody as described in Methods. IP and Output were analyzed by immunoblotting using the anti-D2 antibody. IP and Output were also analyzed using anti-A2A antibody. A representative blot (of three) is shown. A2A immunoreactive bands were quantified and the data were reported in the graph. Values are means ± SEM ( n = 3).
    Rabbit Anti A2a, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "A2A-D2 Heteromers on Striatal Astrocytes: Biochemical and Biophysical Evidence"

    Article Title: A2A-D2 Heteromers on Striatal Astrocytes: Biochemical and Biophysical Evidence

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms20102457

    Interaction between A2A and D2 on rat striatal astrocyte processes: co-immunoprecipitation. ( A ) Aliquots (300 µg) of Triton X-100-soluble proteins obtained from gliosomes were immunoprecipitated with 1 µg of anti-A2A antibody as described in Methods. Immunoprecipitated (IP) and not immunoprecipitated (Output) materials were analyzed by immunoblotting using the anti-A2A antibody. IP and Output were also analyzed using anti-D2 antibody. A representative blot (of three) is shown. D2 immunoreactive bands were quantified and the data were reported in the graph. Values are means ± SEM ( n = 3). ( B ) Aliquots (300 µg) of Triton X-100-soluble proteins obtained from gliosomes were immunoprecipitated with 1 µg of anti-D2 antibody as described in Methods. IP and Output were analyzed by immunoblotting using the anti-D2 antibody. IP and Output were also analyzed using anti-A2A antibody. A representative blot (of three) is shown. A2A immunoreactive bands were quantified and the data were reported in the graph. Values are means ± SEM ( n = 3).
    Figure Legend Snippet: Interaction between A2A and D2 on rat striatal astrocyte processes: co-immunoprecipitation. ( A ) Aliquots (300 µg) of Triton X-100-soluble proteins obtained from gliosomes were immunoprecipitated with 1 µg of anti-A2A antibody as described in Methods. Immunoprecipitated (IP) and not immunoprecipitated (Output) materials were analyzed by immunoblotting using the anti-A2A antibody. IP and Output were also analyzed using anti-D2 antibody. A representative blot (of three) is shown. D2 immunoreactive bands were quantified and the data were reported in the graph. Values are means ± SEM ( n = 3). ( B ) Aliquots (300 µg) of Triton X-100-soluble proteins obtained from gliosomes were immunoprecipitated with 1 µg of anti-D2 antibody as described in Methods. IP and Output were analyzed by immunoblotting using the anti-D2 antibody. IP and Output were also analyzed using anti-A2A antibody. A representative blot (of three) is shown. A2A immunoreactive bands were quantified and the data were reported in the graph. Values are means ± SEM ( n = 3).

    Techniques Used: Immunoprecipitation, Western Blot

    A2A-D2 heterodimers on rat striatal astrocytes: proximity ligation assay. Detection of in situ PLA A2A-D2 heteroreceptor complexes was carried out with primary antibodies (rabbit polyclonal anti-A2AR, mouse monoclonal anti-D2R and goat polyclonal anti-GFAP) in rat striatal slices. ( A ) The merge of the maximum intensity projections of a representative field (240x240 µm; z 10 µm) is shown; GFAP (red), DAPI (blue), PLA for A2A-D2 heteroreceptor complexes appears as yellow clusters. The boxed region (i) is shown at a higher magnification in ( B ). ( C ) Merged confocal images of a single z stack of the boxed region (ii) at a higher magnification: double immunolabeling for GFAP (red) and PLA A2A-D2 heteroreceptor complexes (green). ( D ) Colocalized map of the boxed region (ii); the colocalization analysis plugins—colocalization threshold with GFAP as ROI—was used to create the colocalized map; ImageJ Fiji software. A complete lack of stain for PLA A2A-D2 heteroreceptor complexes was obtained in the negative control experiments, performed avoiding the conjugation of a primary antibody with the Duolink Probes. In the figure the merges of the maximum intensity projections of two representative fields are shown: PLA for A2A-D2. heteroreceptor complexes without the primary anti-A2A antibody ( E ) or the primary anti-D2 antibody ( F ). Scale bar 25 µm or 10 µm
    Figure Legend Snippet: A2A-D2 heterodimers on rat striatal astrocytes: proximity ligation assay. Detection of in situ PLA A2A-D2 heteroreceptor complexes was carried out with primary antibodies (rabbit polyclonal anti-A2AR, mouse monoclonal anti-D2R and goat polyclonal anti-GFAP) in rat striatal slices. ( A ) The merge of the maximum intensity projections of a representative field (240x240 µm; z 10 µm) is shown; GFAP (red), DAPI (blue), PLA for A2A-D2 heteroreceptor complexes appears as yellow clusters. The boxed region (i) is shown at a higher magnification in ( B ). ( C ) Merged confocal images of a single z stack of the boxed region (ii) at a higher magnification: double immunolabeling for GFAP (red) and PLA A2A-D2 heteroreceptor complexes (green). ( D ) Colocalized map of the boxed region (ii); the colocalization analysis plugins—colocalization threshold with GFAP as ROI—was used to create the colocalized map; ImageJ Fiji software. A complete lack of stain for PLA A2A-D2 heteroreceptor complexes was obtained in the negative control experiments, performed avoiding the conjugation of a primary antibody with the Duolink Probes. In the figure the merges of the maximum intensity projections of two representative fields are shown: PLA for A2A-D2. heteroreceptor complexes without the primary anti-A2A antibody ( E ) or the primary anti-D2 antibody ( F ). Scale bar 25 µm or 10 µm

    Techniques Used: Proximity Ligation Assay, In Situ, Immunolabeling, Software, Staining, Negative Control, Conjugation Assay

    a2a  (Alomone Labs)


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    Alomone Labs a2a
    Expression and functional activity of adenosine receptors in A2780, A2780CisR, and HEY cells. a Gene expression of adenosine receptors using RT-PCR. b Protein expression of adenosine receptors using Western blot. c Concentration-dependent effect of adenosine on cAMP-related luminescence in A2780 cells. d–f Concentration-dependent effect of SLV320, PSB603, and SCH 442416 (selective A1, A2B, and <t>A2A</t> antagonists, respectively) on 200 μM adenosine-induced luminescence in A2780 cells
    A2a, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Adenosine enhances cisplatin sensitivity in human ovarian cancer cells"

    Article Title: Adenosine enhances cisplatin sensitivity in human ovarian cancer cells

    Journal: Purinergic Signalling

    doi: 10.1007/s11302-018-9622-7

    Expression and functional activity of adenosine receptors in A2780, A2780CisR, and HEY cells. a Gene expression of adenosine receptors using RT-PCR. b Protein expression of adenosine receptors using Western blot. c Concentration-dependent effect of adenosine on cAMP-related luminescence in A2780 cells. d–f Concentration-dependent effect of SLV320, PSB603, and SCH 442416 (selective A1, A2B, and A2A antagonists, respectively) on 200 μM adenosine-induced luminescence in A2780 cells
    Figure Legend Snippet: Expression and functional activity of adenosine receptors in A2780, A2780CisR, and HEY cells. a Gene expression of adenosine receptors using RT-PCR. b Protein expression of adenosine receptors using Western blot. c Concentration-dependent effect of adenosine on cAMP-related luminescence in A2780 cells. d–f Concentration-dependent effect of SLV320, PSB603, and SCH 442416 (selective A1, A2B, and A2A antagonists, respectively) on 200 μM adenosine-induced luminescence in A2780 cells

    Techniques Used: Expressing, Functional Assay, Activity Assay, Reverse Transcription Polymerase Chain Reaction, Western Blot, Concentration Assay

    igg  (Alomone Labs)


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    Alomone Labs igg
    Igg, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    antibody against a2a  (Alomone Labs)


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    Alomone Labs antibody against a2a
    Antibody Against A2a, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    rabbit anti a2a adenosine receptor antibody  (Alomone Labs)


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    Alomone Labs rabbit anti a2a adenosine receptor antibody
    Western blot (A) and immunofluoresecence assay (IFA) (B) show the presence of <t>A2A</t> adenosine receptor (AR) expression on human primary brain endothelial cells (HBMVEC) and human brain endothelial cell line (HCMEC-D3). For IFA, cells were stained with anti-A2A AR antibody (Green) and anti-human CD31 as endothelial cell marker (Red). Nucleus was counter stained with DAPI (Blue). Scale bar indicates 25 um. (C) FACS analysis (Dot-plot and histogram) shows expression of the extracellular enzymes, CD39 and CD73, in HBMVEC and HCMEC-D3.
    Rabbit Anti A2a Adenosine Receptor Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "A2A adenosine receptor regulates the human blood brain barrier permeability"

    Article Title: A2A adenosine receptor regulates the human blood brain barrier permeability

    Journal: Molecular neurobiology

    doi: 10.1007/s12035-014-8879-2

    Western blot (A) and immunofluoresecence assay (IFA) (B) show the presence of A2A adenosine receptor (AR) expression on human primary brain endothelial cells (HBMVEC) and human brain endothelial cell line (HCMEC-D3). For IFA, cells were stained with anti-A2A AR antibody (Green) and anti-human CD31 as endothelial cell marker (Red). Nucleus was counter stained with DAPI (Blue). Scale bar indicates 25 um. (C) FACS analysis (Dot-plot and histogram) shows expression of the extracellular enzymes, CD39 and CD73, in HBMVEC and HCMEC-D3.
    Figure Legend Snippet: Western blot (A) and immunofluoresecence assay (IFA) (B) show the presence of A2A adenosine receptor (AR) expression on human primary brain endothelial cells (HBMVEC) and human brain endothelial cell line (HCMEC-D3). For IFA, cells were stained with anti-A2A AR antibody (Green) and anti-human CD31 as endothelial cell marker (Red). Nucleus was counter stained with DAPI (Blue). Scale bar indicates 25 um. (C) FACS analysis (Dot-plot and histogram) shows expression of the extracellular enzymes, CD39 and CD73, in HBMVEC and HCMEC-D3.

    Techniques Used: Western Blot, Expressing, Staining, Marker

    (A and B) IFA of VE-cadherin (Green) and F-actin (Red) in HBMVEC cells 5 and 30 minutes post-treatment with Lexiscan and NECA. Additionally, NECA was treated concomitantly with SCH58261 which is A2A specific antagonist. Arrows indicate the disrupted junction formation. Nucleus was counterstained with DAPI (Blue). Scale bar indicates 25 um. (C and D) AR agonists increase the permeability to chemotherapeutics, Gemcitabine, in primary human brain endothelial monolayer. Changes in permeability to the chemotherapeutic drug, Gemcitabine, after AR activation was determined using primary human brain endothelial cell monolayer. Donor chamber was treated with Gemcitabine (Gem) (10 ug/ml) for 5, 15, 30, 60 minutes with or without 1uM of Lexiscan or NECA and donor chambers were removed. Receiver chambers on which the YFP-transfected human glioblastoma cells (U251) were cultured and further incubated for 96 hrs and cell viability was measured by the relative intensity of YFP signal compared to untreated YFP-U251 (n=3). Lexiscan or NECA only treatment group was also set as control to test its effect on glioma cell viability (n=3). Data represents mean ± s.e.m. (* indicates where p<0.05, two tailed student t-test). (E) Selective A2A AR agonist Lexiscan increases the permeability of the BBB to 10 kDa FITC-Dextran in mice. Lexiscan (0.05 mg/kg) was intravenously administered concomitantly with 10 kDa FITC-Dextran and perfused with ice-cold PBS at different time point (n=10). Brain was collected and processed for analysis of FITC-Dextran concentration using fluometry (** indicates where p<0.01, two tailed student t-test). Graph from Figure 2D showing the effect of Lexiscan on hBBB permeability to 10 kDa FITC-Dextran was juxtaposed as inset for comparison. Arrows indicate the time point with maximal FITC-Dextran concentration from two different graphs. (F) Adenosine increases the peremeability of the BBB to 10 kDa FITC-Dextran in mice. Adenosine was intravenously administered three times (0.138 mg/kg, 20 seconds apart) concomitantly with 10 kDa Dextran and perfused with ice cold PBS at 1 minute after treatment (n=2). Brain was collected and processed for analysis of FITC-Dextran concentration using fluometry (* indicates where p<0.05, two tailed student t-test).
    Figure Legend Snippet: (A and B) IFA of VE-cadherin (Green) and F-actin (Red) in HBMVEC cells 5 and 30 minutes post-treatment with Lexiscan and NECA. Additionally, NECA was treated concomitantly with SCH58261 which is A2A specific antagonist. Arrows indicate the disrupted junction formation. Nucleus was counterstained with DAPI (Blue). Scale bar indicates 25 um. (C and D) AR agonists increase the permeability to chemotherapeutics, Gemcitabine, in primary human brain endothelial monolayer. Changes in permeability to the chemotherapeutic drug, Gemcitabine, after AR activation was determined using primary human brain endothelial cell monolayer. Donor chamber was treated with Gemcitabine (Gem) (10 ug/ml) for 5, 15, 30, 60 minutes with or without 1uM of Lexiscan or NECA and donor chambers were removed. Receiver chambers on which the YFP-transfected human glioblastoma cells (U251) were cultured and further incubated for 96 hrs and cell viability was measured by the relative intensity of YFP signal compared to untreated YFP-U251 (n=3). Lexiscan or NECA only treatment group was also set as control to test its effect on glioma cell viability (n=3). Data represents mean ± s.e.m. (* indicates where p<0.05, two tailed student t-test). (E) Selective A2A AR agonist Lexiscan increases the permeability of the BBB to 10 kDa FITC-Dextran in mice. Lexiscan (0.05 mg/kg) was intravenously administered concomitantly with 10 kDa FITC-Dextran and perfused with ice-cold PBS at different time point (n=10). Brain was collected and processed for analysis of FITC-Dextran concentration using fluometry (** indicates where p<0.01, two tailed student t-test). Graph from Figure 2D showing the effect of Lexiscan on hBBB permeability to 10 kDa FITC-Dextran was juxtaposed as inset for comparison. Arrows indicate the time point with maximal FITC-Dextran concentration from two different graphs. (F) Adenosine increases the peremeability of the BBB to 10 kDa FITC-Dextran in mice. Adenosine was intravenously administered three times (0.138 mg/kg, 20 seconds apart) concomitantly with 10 kDa Dextran and perfused with ice cold PBS at 1 minute after treatment (n=2). Brain was collected and processed for analysis of FITC-Dextran concentration using fluometry (* indicates where p<0.05, two tailed student t-test).

    Techniques Used: Permeability, Activation Assay, Transfection, Cell Culture, Incubation, Two Tailed Test, Concentration Assay

    a2ar  (Alomone Labs)


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    Alomone Labs a2ar
    A2ar, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs rabbit anti human monoclonal antibodies against eta
    Immunostaining of sections of stenotic valve from a patient with rheumatic fever. Top left. ET-1 staining associated with fibrotic regions and neovascularization. Top middle. <t>ETA</t> receptor staining of fibrotic region and neovascularization. Top right. H + E-stained section. Lower left. No ET-3 immunostaining. Lower <t>middle.</t> <t>ETB</t> receptor immunostaining of fibrotic region and neovascularization. Lower right. Negative control. ET=endothelin; ETA=endothelin receptor A; ETB=endothelin receptor B
    Rabbit Anti Human Monoclonal Antibodies Against Eta, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs rabbit igg anti p2y12r
    A Immunohistostainings of post-mortem human hemispheric white matter tissue (healthy donor) showing microglial cells (blue, resident microglia, TMEM119) contacting nodes of Ranvier (Na V , brown). B , C Immunofluorescent stainings of post-mortem human hemispheric white matter tissue (healthy donor) showing microglial cells (green; B , resident microglia, TMEM119, and C , homeostatic microglia, <t>P2Y12R)</t> contacting nodes of Ranvier (Na V , red). B ii, C ii 3D reconstructions correspond to the boxed area in B i and C i respectively. Scale bars: (2D) A , B , C 10 μm; (3D), B , C 2 μm. A : n = 6 samples, B , C : n = 5 samples.
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    Alomone Labs anti human anti adenosine a 2a r rabbit polyclonal antibody
    ADORA2A siRNAs decrease ADORA2A mRNA and A <t>2A</t> <t>R</t> protein expression (A) Representation of the structure of the lipid NPs labeled with fluorescent streptavidin and biotinylated targeting antibody and loaded with siRNAs. PE: phosphoethanolamine; PEG: polyethylene glycol (B) Relative quantity (RQ) of ADORA2A mRNA expression in activated HD CD3 + or CD8 + T cells 24 h post-transfection with 10 nM scr or ADORA2A siRNAs (n = 5; 3 CD3 + T cell samples, 2 CD8 + T cell samples). GAPDH (Glyceraldehyde 3-phosphate dehydrogenase) was used as the reference gene. Data are shown as mean ± standard deviation and significance is determined by a paired Student’s t test. (C) (Left) Fold change of normalized A 2A R protein expression in activated HD CD3 + or CD8 + T cells 24 h (n = 5; 3 CD3 + T cell samples, 2 CD8 + T cell samples), 48 h (n = 5; 3 CD3 + T cell samples, 2 CD8 + T cell samples), or 72 h (n = 4; 2 CD3 + T cell samples, 2 CD8 + T cell samples) post-transfection with 10 nM ADORA2A siRNAs as compared to scr-RNAs. (Right) The normalized MFI of A 2A R expression showing the highest degree of A 2A R protein knockdown post-transfection with 10 nM ADORA2A siRNAs as compared to corresponding scr-RNAs for each individual sample regardless of time point at which knockdown occurred. Data are shown as mean ± standard error of the mean (n = 5). Data are compared using a paired Student’s t test. (D) Flow cytometry gating strategy for determining A 2A R expression in transfected cells. The cells were first gated on T lymphocytes and then on GFP fluorescence (indicating that the cells were successfully transfected with siRNAs) to determine A 2A R expression. The unstained sample is indicated in orange in the histogram while the A 2A R expression of cells treated with scr-RNAs is indicated in blue and the A 2A R expression of cells treated with ADORA2A siRNAs is indicated in red. (E) Representative histogram showing A 2A R antibody specificity where CD3 + T cells were stained with A 2A R antibody ± peptide followed by secondary antibody. Unstained cells are indicated in gray, A 2A R antibody + peptide is indicated in red and A 2A R antibody alone is indicated in blue. Shown here is a representative experiment from two identical experiments performed in HDs.
    Anti Human Anti Adenosine A 2a R Rabbit Polyclonal Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs rabbit anti a2a
    Interaction between <t>A2A</t> and D2 on rat striatal astrocyte processes: co-immunoprecipitation. ( A ) Aliquots (300 µg) of Triton X-100-soluble proteins obtained from gliosomes were immunoprecipitated with 1 µg of anti-A2A antibody as described in Methods. Immunoprecipitated (IP) and not immunoprecipitated (Output) materials were analyzed by immunoblotting using the anti-A2A antibody. IP and Output were also analyzed using anti-D2 antibody. A representative blot (of three) is shown. D2 immunoreactive bands were quantified and the data were reported in the graph. Values are means ± SEM ( n = 3). ( B ) Aliquots (300 µg) of Triton X-100-soluble proteins obtained from gliosomes were immunoprecipitated with 1 µg of anti-D2 antibody as described in Methods. IP and Output were analyzed by immunoblotting using the anti-D2 antibody. IP and Output were also analyzed using anti-A2A antibody. A representative blot (of three) is shown. A2A immunoreactive bands were quantified and the data were reported in the graph. Values are means ± SEM ( n = 3).
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    Alomone Labs a2a
    Expression and functional activity of adenosine receptors in A2780, A2780CisR, and HEY cells. a Gene expression of adenosine receptors using RT-PCR. b Protein expression of adenosine receptors using Western blot. c Concentration-dependent effect of adenosine on cAMP-related luminescence in A2780 cells. d–f Concentration-dependent effect of SLV320, PSB603, and SCH 442416 (selective A1, A2B, and <t>A2A</t> antagonists, respectively) on 200 μM adenosine-induced luminescence in A2780 cells
    A2a, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs igg
    Expression and functional activity of adenosine receptors in A2780, A2780CisR, and HEY cells. a Gene expression of adenosine receptors using RT-PCR. b Protein expression of adenosine receptors using Western blot. c Concentration-dependent effect of adenosine on cAMP-related luminescence in A2780 cells. d–f Concentration-dependent effect of SLV320, PSB603, and SCH 442416 (selective A1, A2B, and <t>A2A</t> antagonists, respectively) on 200 μM adenosine-induced luminescence in A2780 cells
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    Alomone Labs antibody against a2a
    Expression and functional activity of adenosine receptors in A2780, A2780CisR, and HEY cells. a Gene expression of adenosine receptors using RT-PCR. b Protein expression of adenosine receptors using Western blot. c Concentration-dependent effect of adenosine on cAMP-related luminescence in A2780 cells. d–f Concentration-dependent effect of SLV320, PSB603, and SCH 442416 (selective A1, A2B, and <t>A2A</t> antagonists, respectively) on 200 μM adenosine-induced luminescence in A2780 cells
    Antibody Against A2a, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs rabbit anti a2a adenosine receptor antibody
    Western blot (A) and immunofluoresecence assay (IFA) (B) show the presence of <t>A2A</t> adenosine receptor (AR) expression on human primary brain endothelial cells (HBMVEC) and human brain endothelial cell line (HCMEC-D3). For IFA, cells were stained with anti-A2A AR antibody (Green) and anti-human CD31 as endothelial cell marker (Red). Nucleus was counter stained with DAPI (Blue). Scale bar indicates 25 um. (C) FACS analysis (Dot-plot and histogram) shows expression of the extracellular enzymes, CD39 and CD73, in HBMVEC and HCMEC-D3.
    Rabbit Anti A2a Adenosine Receptor Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs a2ar
    Western blot (A) and immunofluoresecence assay (IFA) (B) show the presence of <t>A2A</t> adenosine receptor (AR) expression on human primary brain endothelial cells (HBMVEC) and human brain endothelial cell line (HCMEC-D3). For IFA, cells were stained with anti-A2A AR antibody (Green) and anti-human CD31 as endothelial cell marker (Red). Nucleus was counter stained with DAPI (Blue). Scale bar indicates 25 um. (C) FACS analysis (Dot-plot and histogram) shows expression of the extracellular enzymes, CD39 and CD73, in HBMVEC and HCMEC-D3.
    A2ar, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/a2ar/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
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    Image Search Results


    Immunostaining of sections of stenotic valve from a patient with rheumatic fever. Top left. ET-1 staining associated with fibrotic regions and neovascularization. Top middle. ETA receptor staining of fibrotic region and neovascularization. Top right. H + E-stained section. Lower left. No ET-3 immunostaining. Lower middle. ETB receptor immunostaining of fibrotic region and neovascularization. Lower right. Negative control. ET=endothelin; ETA=endothelin receptor A; ETB=endothelin receptor B

    Journal: Revista Brasileira de Cirurgia Cardiovascular : órgão oficial da Sociedade Brasileira de Cirurgia Cardiovascular

    Article Title: Analysis of immunostaining and western blotting of endothelin 1 and its receptors in mitral stenosis

    doi: 10.5935/1678-9741.20150004

    Figure Lengend Snippet: Immunostaining of sections of stenotic valve from a patient with rheumatic fever. Top left. ET-1 staining associated with fibrotic regions and neovascularization. Top middle. ETA receptor staining of fibrotic region and neovascularization. Top right. H + E-stained section. Lower left. No ET-3 immunostaining. Lower middle. ETB receptor immunostaining of fibrotic region and neovascularization. Lower right. Negative control. ET=endothelin; ETA=endothelin receptor A; ETB=endothelin receptor B

    Article Snippet: Antibodies and their dilution: Mouse anti-human monoclonal antibody against endothelial cells (CD31, clone JC70A, 1:100, DakoCytomation, Denmark); rabbit anti-human monoclonal antibody against ET-1 and ET-3 (1:250, R & D Labs), rabbit anti-human monoclonal antibodies against ETA and ETB receptors (both 1:200, Alomone Labs, Israel) and AS02/Thy-1 for fibroblasts (1:200, DakoCytomation, Denmark).

    Techniques: Immunostaining, Staining, Negative Control

    A Immunohistostainings of post-mortem human hemispheric white matter tissue (healthy donor) showing microglial cells (blue, resident microglia, TMEM119) contacting nodes of Ranvier (Na V , brown). B , C Immunofluorescent stainings of post-mortem human hemispheric white matter tissue (healthy donor) showing microglial cells (green; B , resident microglia, TMEM119, and C , homeostatic microglia, P2Y12R) contacting nodes of Ranvier (Na V , red). B ii, C ii 3D reconstructions correspond to the boxed area in B i and C i respectively. Scale bars: (2D) A , B , C 10 μm; (3D), B , C 2 μm. A : n = 6 samples, B , C : n = 5 samples.

    Journal: Nature Communications

    Article Title: Microglia-neuron interaction at nodes of Ranvier depends on neuronal activity through potassium release and contributes to remyelination

    doi: 10.1038/s41467-021-25486-7

    Figure Lengend Snippet: A Immunohistostainings of post-mortem human hemispheric white matter tissue (healthy donor) showing microglial cells (blue, resident microglia, TMEM119) contacting nodes of Ranvier (Na V , brown). B , C Immunofluorescent stainings of post-mortem human hemispheric white matter tissue (healthy donor) showing microglial cells (green; B , resident microglia, TMEM119, and C , homeostatic microglia, P2Y12R) contacting nodes of Ranvier (Na V , red). B ii, C ii 3D reconstructions correspond to the boxed area in B i and C i respectively. Scale bars: (2D) A , B , C 10 μm; (3D), B , C 2 μm. A : n = 6 samples, B , C : n = 5 samples.

    Article Snippet: Primary antibodies: mouse IgG2a anti-AnkyrinG (clone N106/36; 1:100), mouse IgG2b anti-AnkyrinG (clone N106/65; 1:75), and mouse IgG1 anti-Caspr (1:100), all from Neuromab; mouse IgG1 anti-Pan Na v (clone K58/35; 1:150; Sigma); mouse anti-Calbindin (1:500; Sigma), rabbit anti-Calbindin (1:300; Swant), rabbit anti-Caspr (1:300; Abcam), rat anti-PLP (1:10; kindly provided by Dr. K. Ikenaka, Okasaki, Japan), mouse IgG2b anti-MBP (1:200; SMI99, Sigma), rabbit IgG anti-Iba1 (1:500; Wako), chicken anti-GFAP (1:500; Aves Labs), rat anti-PDGFrα (1:100; BD Biosciences), rabbit IgG anti-TMEM119 (1:100; Sigma), rabbit IgG anti-P2Y12r (1:300; Alomone, human tissue), rabbit anti-P2Y12R (1:300; Anaspec, mouse tissue), chicken anti-GFP (1:250; Millipore), mouse IgG2a anti-iNOS (1:100; BD Biosciences), goat anti-IGF1 (1:50; R&D System), and chicken anti-mCherry (1:1000; Abcam).

    Techniques:

    ADORA2A siRNAs decrease ADORA2A mRNA and A 2A R protein expression (A) Representation of the structure of the lipid NPs labeled with fluorescent streptavidin and biotinylated targeting antibody and loaded with siRNAs. PE: phosphoethanolamine; PEG: polyethylene glycol (B) Relative quantity (RQ) of ADORA2A mRNA expression in activated HD CD3 + or CD8 + T cells 24 h post-transfection with 10 nM scr or ADORA2A siRNAs (n = 5; 3 CD3 + T cell samples, 2 CD8 + T cell samples). GAPDH (Glyceraldehyde 3-phosphate dehydrogenase) was used as the reference gene. Data are shown as mean ± standard deviation and significance is determined by a paired Student’s t test. (C) (Left) Fold change of normalized A 2A R protein expression in activated HD CD3 + or CD8 + T cells 24 h (n = 5; 3 CD3 + T cell samples, 2 CD8 + T cell samples), 48 h (n = 5; 3 CD3 + T cell samples, 2 CD8 + T cell samples), or 72 h (n = 4; 2 CD3 + T cell samples, 2 CD8 + T cell samples) post-transfection with 10 nM ADORA2A siRNAs as compared to scr-RNAs. (Right) The normalized MFI of A 2A R expression showing the highest degree of A 2A R protein knockdown post-transfection with 10 nM ADORA2A siRNAs as compared to corresponding scr-RNAs for each individual sample regardless of time point at which knockdown occurred. Data are shown as mean ± standard error of the mean (n = 5). Data are compared using a paired Student’s t test. (D) Flow cytometry gating strategy for determining A 2A R expression in transfected cells. The cells were first gated on T lymphocytes and then on GFP fluorescence (indicating that the cells were successfully transfected with siRNAs) to determine A 2A R expression. The unstained sample is indicated in orange in the histogram while the A 2A R expression of cells treated with scr-RNAs is indicated in blue and the A 2A R expression of cells treated with ADORA2A siRNAs is indicated in red. (E) Representative histogram showing A 2A R antibody specificity where CD3 + T cells were stained with A 2A R antibody ± peptide followed by secondary antibody. Unstained cells are indicated in gray, A 2A R antibody + peptide is indicated in red and A 2A R antibody alone is indicated in blue. Shown here is a representative experiment from two identical experiments performed in HDs.

    Journal: Molecular Therapy. Methods & Clinical Development

    Article Title: Targeted knockdown of the adenosine A 2A receptor by lipid NPs rescues the chemotaxis of head and neck cancer memory T cells

    doi: 10.1016/j.omtm.2021.03.001

    Figure Lengend Snippet: ADORA2A siRNAs decrease ADORA2A mRNA and A 2A R protein expression (A) Representation of the structure of the lipid NPs labeled with fluorescent streptavidin and biotinylated targeting antibody and loaded with siRNAs. PE: phosphoethanolamine; PEG: polyethylene glycol (B) Relative quantity (RQ) of ADORA2A mRNA expression in activated HD CD3 + or CD8 + T cells 24 h post-transfection with 10 nM scr or ADORA2A siRNAs (n = 5; 3 CD3 + T cell samples, 2 CD8 + T cell samples). GAPDH (Glyceraldehyde 3-phosphate dehydrogenase) was used as the reference gene. Data are shown as mean ± standard deviation and significance is determined by a paired Student’s t test. (C) (Left) Fold change of normalized A 2A R protein expression in activated HD CD3 + or CD8 + T cells 24 h (n = 5; 3 CD3 + T cell samples, 2 CD8 + T cell samples), 48 h (n = 5; 3 CD3 + T cell samples, 2 CD8 + T cell samples), or 72 h (n = 4; 2 CD3 + T cell samples, 2 CD8 + T cell samples) post-transfection with 10 nM ADORA2A siRNAs as compared to scr-RNAs. (Right) The normalized MFI of A 2A R expression showing the highest degree of A 2A R protein knockdown post-transfection with 10 nM ADORA2A siRNAs as compared to corresponding scr-RNAs for each individual sample regardless of time point at which knockdown occurred. Data are shown as mean ± standard error of the mean (n = 5). Data are compared using a paired Student’s t test. (D) Flow cytometry gating strategy for determining A 2A R expression in transfected cells. The cells were first gated on T lymphocytes and then on GFP fluorescence (indicating that the cells were successfully transfected with siRNAs) to determine A 2A R expression. The unstained sample is indicated in orange in the histogram while the A 2A R expression of cells treated with scr-RNAs is indicated in blue and the A 2A R expression of cells treated with ADORA2A siRNAs is indicated in red. (E) Representative histogram showing A 2A R antibody specificity where CD3 + T cells were stained with A 2A R antibody ± peptide followed by secondary antibody. Unstained cells are indicated in gray, A 2A R antibody + peptide is indicated in red and A 2A R antibody alone is indicated in blue. Shown here is a representative experiment from two identical experiments performed in HDs.

    Article Snippet: Anti-human anti-adenosine A 2A R rabbit polyclonal antibody (0.75 μg) was incubated with 1.5 μg A 2A R blocking peptide (Alomone) at a 1:2 ratio (1.5 μg peptide was added to 0.75 μg antibody).

    Techniques: Expressing, Labeling, Transfection, Standard Deviation, Flow Cytometry, Fluorescence, Staining

    CD45RO(A 2A R)-NPs rescue the chemotactic ability of HNSCC CD8 + memory T cells in the presence of adenosine (A and B) Representative two point trajectories of T cells migrating along the CXCL10 gradient (green) or CXCL10 + adenosine (ADO) gradient (blue) in HNSCC CD8 + memory T cells treated with (A) CD45RO(scr)-NPs or (B) CD45RO(A 2A R)-NPs. Trajectories artificially set to start at the origin with the red triangle representing the Y-COM. (C) Y-COM of HNSCC CD8 + memory T cells treated with (left) CD45RO(scr)-NPs (n = 6) or (right) CD45RO(A 2A R)-NPs (n = 6) in the presence of CXCL10 or CXCL10 + adenosine. Significance was determined using (left) a Wilcoxon signed rank test or (right) paired Student’s t test. (D) Percent inhibition of Y-COM of HNSCC CD8 + memory T cells in the presence of CXCL10 + adenosine compared to CXCL10 alone when treated with CD45RO(scr)-NPs (n = 6) compared to CD45RO(A 2A R)-NPs (n = 6). Data are represented single dot plots (colored dots; wherein each dot represents a single donor) and as mean ± standard deviation (shown in black). Significance was determined using Student’s t test.

    Journal: Molecular Therapy. Methods & Clinical Development

    Article Title: Targeted knockdown of the adenosine A 2A receptor by lipid NPs rescues the chemotaxis of head and neck cancer memory T cells

    doi: 10.1016/j.omtm.2021.03.001

    Figure Lengend Snippet: CD45RO(A 2A R)-NPs rescue the chemotactic ability of HNSCC CD8 + memory T cells in the presence of adenosine (A and B) Representative two point trajectories of T cells migrating along the CXCL10 gradient (green) or CXCL10 + adenosine (ADO) gradient (blue) in HNSCC CD8 + memory T cells treated with (A) CD45RO(scr)-NPs or (B) CD45RO(A 2A R)-NPs. Trajectories artificially set to start at the origin with the red triangle representing the Y-COM. (C) Y-COM of HNSCC CD8 + memory T cells treated with (left) CD45RO(scr)-NPs (n = 6) or (right) CD45RO(A 2A R)-NPs (n = 6) in the presence of CXCL10 or CXCL10 + adenosine. Significance was determined using (left) a Wilcoxon signed rank test or (right) paired Student’s t test. (D) Percent inhibition of Y-COM of HNSCC CD8 + memory T cells in the presence of CXCL10 + adenosine compared to CXCL10 alone when treated with CD45RO(scr)-NPs (n = 6) compared to CD45RO(A 2A R)-NPs (n = 6). Data are represented single dot plots (colored dots; wherein each dot represents a single donor) and as mean ± standard deviation (shown in black). Significance was determined using Student’s t test.

    Article Snippet: Anti-human anti-adenosine A 2A R rabbit polyclonal antibody (0.75 μg) was incubated with 1.5 μg A 2A R blocking peptide (Alomone) at a 1:2 ratio (1.5 μg peptide was added to 0.75 μg antibody).

    Techniques: Inhibition, Standard Deviation

    Interaction between A2A and D2 on rat striatal astrocyte processes: co-immunoprecipitation. ( A ) Aliquots (300 µg) of Triton X-100-soluble proteins obtained from gliosomes were immunoprecipitated with 1 µg of anti-A2A antibody as described in Methods. Immunoprecipitated (IP) and not immunoprecipitated (Output) materials were analyzed by immunoblotting using the anti-A2A antibody. IP and Output were also analyzed using anti-D2 antibody. A representative blot (of three) is shown. D2 immunoreactive bands were quantified and the data were reported in the graph. Values are means ± SEM ( n = 3). ( B ) Aliquots (300 µg) of Triton X-100-soluble proteins obtained from gliosomes were immunoprecipitated with 1 µg of anti-D2 antibody as described in Methods. IP and Output were analyzed by immunoblotting using the anti-D2 antibody. IP and Output were also analyzed using anti-A2A antibody. A representative blot (of three) is shown. A2A immunoreactive bands were quantified and the data were reported in the graph. Values are means ± SEM ( n = 3).

    Journal: International Journal of Molecular Sciences

    Article Title: A2A-D2 Heteromers on Striatal Astrocytes: Biochemical and Biophysical Evidence

    doi: 10.3390/ijms20102457

    Figure Lengend Snippet: Interaction between A2A and D2 on rat striatal astrocyte processes: co-immunoprecipitation. ( A ) Aliquots (300 µg) of Triton X-100-soluble proteins obtained from gliosomes were immunoprecipitated with 1 µg of anti-A2A antibody as described in Methods. Immunoprecipitated (IP) and not immunoprecipitated (Output) materials were analyzed by immunoblotting using the anti-A2A antibody. IP and Output were also analyzed using anti-D2 antibody. A representative blot (of three) is shown. D2 immunoreactive bands were quantified and the data were reported in the graph. Values are means ± SEM ( n = 3). ( B ) Aliquots (300 µg) of Triton X-100-soluble proteins obtained from gliosomes were immunoprecipitated with 1 µg of anti-D2 antibody as described in Methods. IP and Output were analyzed by immunoblotting using the anti-D2 antibody. IP and Output were also analyzed using anti-A2A antibody. A representative blot (of three) is shown. A2A immunoreactive bands were quantified and the data were reported in the graph. Values are means ± SEM ( n = 3).

    Article Snippet: Mouse anti-A2A primary antibody was purchased from Merck Millipore Corporation (05-717; [ ]); goat anti-GFAP primary antibody was purchased from Santa Cruz Biotechnoloy Inc; rabbit anti-A2A and anti-D2 primary antibodies were purchased from Alomone Labs.

    Techniques: Immunoprecipitation, Western Blot

    A2A-D2 heterodimers on rat striatal astrocytes: proximity ligation assay. Detection of in situ PLA A2A-D2 heteroreceptor complexes was carried out with primary antibodies (rabbit polyclonal anti-A2AR, mouse monoclonal anti-D2R and goat polyclonal anti-GFAP) in rat striatal slices. ( A ) The merge of the maximum intensity projections of a representative field (240x240 µm; z 10 µm) is shown; GFAP (red), DAPI (blue), PLA for A2A-D2 heteroreceptor complexes appears as yellow clusters. The boxed region (i) is shown at a higher magnification in ( B ). ( C ) Merged confocal images of a single z stack of the boxed region (ii) at a higher magnification: double immunolabeling for GFAP (red) and PLA A2A-D2 heteroreceptor complexes (green). ( D ) Colocalized map of the boxed region (ii); the colocalization analysis plugins—colocalization threshold with GFAP as ROI—was used to create the colocalized map; ImageJ Fiji software. A complete lack of stain for PLA A2A-D2 heteroreceptor complexes was obtained in the negative control experiments, performed avoiding the conjugation of a primary antibody with the Duolink Probes. In the figure the merges of the maximum intensity projections of two representative fields are shown: PLA for A2A-D2. heteroreceptor complexes without the primary anti-A2A antibody ( E ) or the primary anti-D2 antibody ( F ). Scale bar 25 µm or 10 µm

    Journal: International Journal of Molecular Sciences

    Article Title: A2A-D2 Heteromers on Striatal Astrocytes: Biochemical and Biophysical Evidence

    doi: 10.3390/ijms20102457

    Figure Lengend Snippet: A2A-D2 heterodimers on rat striatal astrocytes: proximity ligation assay. Detection of in situ PLA A2A-D2 heteroreceptor complexes was carried out with primary antibodies (rabbit polyclonal anti-A2AR, mouse monoclonal anti-D2R and goat polyclonal anti-GFAP) in rat striatal slices. ( A ) The merge of the maximum intensity projections of a representative field (240x240 µm; z 10 µm) is shown; GFAP (red), DAPI (blue), PLA for A2A-D2 heteroreceptor complexes appears as yellow clusters. The boxed region (i) is shown at a higher magnification in ( B ). ( C ) Merged confocal images of a single z stack of the boxed region (ii) at a higher magnification: double immunolabeling for GFAP (red) and PLA A2A-D2 heteroreceptor complexes (green). ( D ) Colocalized map of the boxed region (ii); the colocalization analysis plugins—colocalization threshold with GFAP as ROI—was used to create the colocalized map; ImageJ Fiji software. A complete lack of stain for PLA A2A-D2 heteroreceptor complexes was obtained in the negative control experiments, performed avoiding the conjugation of a primary antibody with the Duolink Probes. In the figure the merges of the maximum intensity projections of two representative fields are shown: PLA for A2A-D2. heteroreceptor complexes without the primary anti-A2A antibody ( E ) or the primary anti-D2 antibody ( F ). Scale bar 25 µm or 10 µm

    Article Snippet: Mouse anti-A2A primary antibody was purchased from Merck Millipore Corporation (05-717; [ ]); goat anti-GFAP primary antibody was purchased from Santa Cruz Biotechnoloy Inc; rabbit anti-A2A and anti-D2 primary antibodies were purchased from Alomone Labs.

    Techniques: Proximity Ligation Assay, In Situ, Immunolabeling, Software, Staining, Negative Control, Conjugation Assay

    Expression and functional activity of adenosine receptors in A2780, A2780CisR, and HEY cells. a Gene expression of adenosine receptors using RT-PCR. b Protein expression of adenosine receptors using Western blot. c Concentration-dependent effect of adenosine on cAMP-related luminescence in A2780 cells. d–f Concentration-dependent effect of SLV320, PSB603, and SCH 442416 (selective A1, A2B, and A2A antagonists, respectively) on 200 μM adenosine-induced luminescence in A2780 cells

    Journal: Purinergic Signalling

    Article Title: Adenosine enhances cisplatin sensitivity in human ovarian cancer cells

    doi: 10.1007/s11302-018-9622-7

    Figure Lengend Snippet: Expression and functional activity of adenosine receptors in A2780, A2780CisR, and HEY cells. a Gene expression of adenosine receptors using RT-PCR. b Protein expression of adenosine receptors using Western blot. c Concentration-dependent effect of adenosine on cAMP-related luminescence in A2780 cells. d–f Concentration-dependent effect of SLV320, PSB603, and SCH 442416 (selective A1, A2B, and A2A antagonists, respectively) on 200 μM adenosine-induced luminescence in A2780 cells

    Article Snippet: Rabbit anti-A1, A2A, A2B, and A3 adenosine receptor antibodies were obtained from Alomone labs (Israel).

    Techniques: Expressing, Functional Assay, Activity Assay, Reverse Transcription Polymerase Chain Reaction, Western Blot, Concentration Assay

    Western blot (A) and immunofluoresecence assay (IFA) (B) show the presence of A2A adenosine receptor (AR) expression on human primary brain endothelial cells (HBMVEC) and human brain endothelial cell line (HCMEC-D3). For IFA, cells were stained with anti-A2A AR antibody (Green) and anti-human CD31 as endothelial cell marker (Red). Nucleus was counter stained with DAPI (Blue). Scale bar indicates 25 um. (C) FACS analysis (Dot-plot and histogram) shows expression of the extracellular enzymes, CD39 and CD73, in HBMVEC and HCMEC-D3.

    Journal: Molecular neurobiology

    Article Title: A2A adenosine receptor regulates the human blood brain barrier permeability

    doi: 10.1007/s12035-014-8879-2

    Figure Lengend Snippet: Western blot (A) and immunofluoresecence assay (IFA) (B) show the presence of A2A adenosine receptor (AR) expression on human primary brain endothelial cells (HBMVEC) and human brain endothelial cell line (HCMEC-D3). For IFA, cells were stained with anti-A2A AR antibody (Green) and anti-human CD31 as endothelial cell marker (Red). Nucleus was counter stained with DAPI (Blue). Scale bar indicates 25 um. (C) FACS analysis (Dot-plot and histogram) shows expression of the extracellular enzymes, CD39 and CD73, in HBMVEC and HCMEC-D3.

    Article Snippet: Rabbit anti-A2A adenosine receptor antibody was purchased from Alomone labs (Jerusalem, Israel).

    Techniques: Western Blot, Expressing, Staining, Marker

    (A and B) IFA of VE-cadherin (Green) and F-actin (Red) in HBMVEC cells 5 and 30 minutes post-treatment with Lexiscan and NECA. Additionally, NECA was treated concomitantly with SCH58261 which is A2A specific antagonist. Arrows indicate the disrupted junction formation. Nucleus was counterstained with DAPI (Blue). Scale bar indicates 25 um. (C and D) AR agonists increase the permeability to chemotherapeutics, Gemcitabine, in primary human brain endothelial monolayer. Changes in permeability to the chemotherapeutic drug, Gemcitabine, after AR activation was determined using primary human brain endothelial cell monolayer. Donor chamber was treated with Gemcitabine (Gem) (10 ug/ml) for 5, 15, 30, 60 minutes with or without 1uM of Lexiscan or NECA and donor chambers were removed. Receiver chambers on which the YFP-transfected human glioblastoma cells (U251) were cultured and further incubated for 96 hrs and cell viability was measured by the relative intensity of YFP signal compared to untreated YFP-U251 (n=3). Lexiscan or NECA only treatment group was also set as control to test its effect on glioma cell viability (n=3). Data represents mean ± s.e.m. (* indicates where p<0.05, two tailed student t-test). (E) Selective A2A AR agonist Lexiscan increases the permeability of the BBB to 10 kDa FITC-Dextran in mice. Lexiscan (0.05 mg/kg) was intravenously administered concomitantly with 10 kDa FITC-Dextran and perfused with ice-cold PBS at different time point (n=10). Brain was collected and processed for analysis of FITC-Dextran concentration using fluometry (** indicates where p<0.01, two tailed student t-test). Graph from Figure 2D showing the effect of Lexiscan on hBBB permeability to 10 kDa FITC-Dextran was juxtaposed as inset for comparison. Arrows indicate the time point with maximal FITC-Dextran concentration from two different graphs. (F) Adenosine increases the peremeability of the BBB to 10 kDa FITC-Dextran in mice. Adenosine was intravenously administered three times (0.138 mg/kg, 20 seconds apart) concomitantly with 10 kDa Dextran and perfused with ice cold PBS at 1 minute after treatment (n=2). Brain was collected and processed for analysis of FITC-Dextran concentration using fluometry (* indicates where p<0.05, two tailed student t-test).

    Journal: Molecular neurobiology

    Article Title: A2A adenosine receptor regulates the human blood brain barrier permeability

    doi: 10.1007/s12035-014-8879-2

    Figure Lengend Snippet: (A and B) IFA of VE-cadherin (Green) and F-actin (Red) in HBMVEC cells 5 and 30 minutes post-treatment with Lexiscan and NECA. Additionally, NECA was treated concomitantly with SCH58261 which is A2A specific antagonist. Arrows indicate the disrupted junction formation. Nucleus was counterstained with DAPI (Blue). Scale bar indicates 25 um. (C and D) AR agonists increase the permeability to chemotherapeutics, Gemcitabine, in primary human brain endothelial monolayer. Changes in permeability to the chemotherapeutic drug, Gemcitabine, after AR activation was determined using primary human brain endothelial cell monolayer. Donor chamber was treated with Gemcitabine (Gem) (10 ug/ml) for 5, 15, 30, 60 minutes with or without 1uM of Lexiscan or NECA and donor chambers were removed. Receiver chambers on which the YFP-transfected human glioblastoma cells (U251) were cultured and further incubated for 96 hrs and cell viability was measured by the relative intensity of YFP signal compared to untreated YFP-U251 (n=3). Lexiscan or NECA only treatment group was also set as control to test its effect on glioma cell viability (n=3). Data represents mean ± s.e.m. (* indicates where p<0.05, two tailed student t-test). (E) Selective A2A AR agonist Lexiscan increases the permeability of the BBB to 10 kDa FITC-Dextran in mice. Lexiscan (0.05 mg/kg) was intravenously administered concomitantly with 10 kDa FITC-Dextran and perfused with ice-cold PBS at different time point (n=10). Brain was collected and processed for analysis of FITC-Dextran concentration using fluometry (** indicates where p<0.01, two tailed student t-test). Graph from Figure 2D showing the effect of Lexiscan on hBBB permeability to 10 kDa FITC-Dextran was juxtaposed as inset for comparison. Arrows indicate the time point with maximal FITC-Dextran concentration from two different graphs. (F) Adenosine increases the peremeability of the BBB to 10 kDa FITC-Dextran in mice. Adenosine was intravenously administered three times (0.138 mg/kg, 20 seconds apart) concomitantly with 10 kDa Dextran and perfused with ice cold PBS at 1 minute after treatment (n=2). Brain was collected and processed for analysis of FITC-Dextran concentration using fluometry (* indicates where p<0.05, two tailed student t-test).

    Article Snippet: Rabbit anti-A2A adenosine receptor antibody was purchased from Alomone labs (Jerusalem, Israel).

    Techniques: Permeability, Activation Assay, Transfection, Cell Culture, Incubation, Two Tailed Test, Concentration Assay