p2y receptor antibodies  (Alomone Labs)


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    Structured Review

    Alomone Labs p2y receptor antibodies
    Confocal images of examples of double immunofluorescence labeling to characterize the colocalization of P2X 1,2,4,7 and <t>P2Y</t> 1,6,12 receptor subtypes on a–f OX42- or g–o GSA-B4-labeled microglial cells in the NAc of rats 4 days after stab-wound injury ( thin arrow process-bearing microglial cell; thick arrow activated microglial cell) ( scale bars : a, b = 20 μm; c, d = 100 μm; e–i = 20 μm; j, k = 100 μm; l, m = 20 μm; n, o = 50 μm)
    P2y Receptor Antibodies, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p2y receptor antibodies/product/Alomone Labs
    Average 85 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    p2y receptor antibodies - by Bioz Stars, 2022-08
    85/100 stars

    Images

    1) Product Images from "Involvement of P2X and P2Y receptors in microglial activation in vivo"

    Article Title: Involvement of P2X and P2Y receptors in microglial activation in vivo

    Journal: Purinergic Signalling

    doi: 10.1007/s11302-007-9082-y

    Confocal images of examples of double immunofluorescence labeling to characterize the colocalization of P2X 1,2,4,7 and P2Y 1,6,12 receptor subtypes on a–f OX42- or g–o GSA-B4-labeled microglial cells in the NAc of rats 4 days after stab-wound injury ( thin arrow process-bearing microglial cell; thick arrow activated microglial cell) ( scale bars : a, b = 20 μm; c, d = 100 μm; e–i = 20 μm; j, k = 100 μm; l, m = 20 μm; n, o = 50 μm)
    Figure Legend Snippet: Confocal images of examples of double immunofluorescence labeling to characterize the colocalization of P2X 1,2,4,7 and P2Y 1,6,12 receptor subtypes on a–f OX42- or g–o GSA-B4-labeled microglial cells in the NAc of rats 4 days after stab-wound injury ( thin arrow process-bearing microglial cell; thick arrow activated microglial cell) ( scale bars : a, b = 20 μm; c, d = 100 μm; e–i = 20 μm; j, k = 100 μm; l, m = 20 μm; n, o = 50 μm)

    Techniques Used: Immunofluorescence, Labeling

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    Alomone Labs p2y6 receptors
    RT-PCR and sequence analysis of porcine P2Y2 , P2Y4 and <t>P2Y6</t> receptors
    P2y6 Receptors, 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/p2y6 receptors/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p2y6 receptors - by Bioz Stars, 2022-08
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    93
    Alomone Labs p2y2
    RT-PCR and sequence analysis of porcine <t>P2Y2</t> , P2Y4 and P2Y6 receptors
    P2y2, 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/p2y2/product/Alomone Labs
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    93
    Alomone Labs p2y4
    Subcellular distribution of P2Y2, <t>P2Y4,</t> and P2Y6 receptors in MEFs. WT or KI MEFs were cultured and, after changing the medium, were maintained in the absence or presence of 1 μ M DFU for 2 h. Cells were fixed with paraformaldehyde (4%; pH 7.2) and permeabilized with cold methanol at RT. After incubation with anti-P2Y2, anti-P2Y4 and anti-P2Y6 antibodies (1 : 500) overnight at 4°C, cells were visualized by confocal microscopy using a FITC-conjugated secondary Ab (Alexa-Fluor 488, 1 : 1000). Nuclei were stained with Hoechst 33258. Coverslips were mounted in Prolong Gold antifade reagent (Molecular Probes) and the intensity of the fluorescence was measured using Image J software (NIH, Bethesda, MD, USA). Results show the mean + SD of three experiments. ** P
    P2y4, 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/p2y4/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    Image Search Results


    RT-PCR and sequence analysis of porcine P2Y2 , P2Y4 and P2Y6 receptors

    Journal:

    Article Title: Evidence for the Expression of Multiple Uracil Nucleotide-Stimulated P2 Receptors Coupled to Smooth Muscle Contraction in Porcine Isolated Arteries

    doi: 10.1038/sj.bjp.0707120

    Figure Lengend Snippet: RT-PCR and sequence analysis of porcine P2Y2 , P2Y4 and P2Y6 receptors

    Article Snippet: For example, reports describe expression of mRNA for P2Y2 , P2Y4 and P2Y6 receptors in human coronary artery , P2Y2 and P2Y6 receptors in human omental and cerebral arteries ( ) and P2Y2 , P2Y4 and P2Y6 receptors in human placental vasculature ( ).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Sequencing

    P2Y6 receptor expressed on microglia and dramatically increased within 3 d after tMCAO. A, Western blot and the quantification of P2Y6 receptor at 0 h, 6 h, 12 h, 1, and 3 d after ischemic stroke. B, Quantitative of RT‐PCR analysis for the expression of P2Y6 receptor at 0 h, 6 h, 12 h, 1, and 3 d after tMCAO. C, Representative fluorescence images of P2Y6 receptor in the contralateral and ipsilateral hemispheres of mice after tMCAO. Scale bar = 100 μm. D, Z‐stack for representative fluorescence images of (a) P2Y6 receptor (green) and Iba1 (red), (b) P2Y6 receptor (green) and GFAP (red), and (c) P2Y6 receptor (green) and Tuj 1 (red). Scale bar = 10 μm. Low magnification images are also presented respectively. Scale bar = 50 μm. Data were presented as mean ± SEM (n = 3 per group). * P

    Journal: CNS Neuroscience & Therapeutics

    Article Title: P2Y6 receptor inhibition aggravates ischemic brain injury by reducing microglial phagocytosis, et al. P2Y6 receptor inhibition aggravates ischemic brain injury by reducing microglial phagocytosis

    doi: 10.1111/cns.13296

    Figure Lengend Snippet: P2Y6 receptor expressed on microglia and dramatically increased within 3 d after tMCAO. A, Western blot and the quantification of P2Y6 receptor at 0 h, 6 h, 12 h, 1, and 3 d after ischemic stroke. B, Quantitative of RT‐PCR analysis for the expression of P2Y6 receptor at 0 h, 6 h, 12 h, 1, and 3 d after tMCAO. C, Representative fluorescence images of P2Y6 receptor in the contralateral and ipsilateral hemispheres of mice after tMCAO. Scale bar = 100 μm. D, Z‐stack for representative fluorescence images of (a) P2Y6 receptor (green) and Iba1 (red), (b) P2Y6 receptor (green) and GFAP (red), and (c) P2Y6 receptor (green) and Tuj 1 (red). Scale bar = 10 μm. Low magnification images are also presented respectively. Scale bar = 50 μm. Data were presented as mean ± SEM (n = 3 per group). * P

    Article Snippet: Then, the sections were incubated with primary antibodies anti‐P2Y6 receptor (1:50, Alomone), Iba1 (1:200, Novusbio), GFAP (1:200, Millipore), Tuj‐1 (1:200, Millipore), NeuN (1:200, Millipore), or Lamp‐2 (1:200, Millipore) overnight at 4°C.

    Techniques: Western Blot, Quantitative RT-PCR, Expressing, Fluorescence, Mouse Assay

    P2Y6 receptor antagonist MRS2578 inhibit MLCK expression in mice at 3 d after tMCAO and cultured microglia. A, (a) Representative fluorescence images of MLCK in Iba‐1 + cultured primary microglia. Scale bar = 25 μm. (b) Quantitative analysis of MLCK in each Iba1 + microglia. n = 4 per group. B, (a) Western blot for the expression of MLCK in the control, LPS+UDP and LPD+UDP+MRS2578 group. (b) Quantification for the intensity ratios of MLCK/β‐actin in the control, LPS+UDP and LPD+UDP+MRS2578 group. C, (a) Western blot for the expression of MLCK in the sham, saline, and MRS2578 group at 3 d after tMCAO. (b) Quantification for the intensity ratios of MLCK/β‐actin in the sham, saline, and MRS2578 group. Data were presented as mean ± SEM. n = 3‐4 per group. * P

    Journal: CNS Neuroscience & Therapeutics

    Article Title: P2Y6 receptor inhibition aggravates ischemic brain injury by reducing microglial phagocytosis, et al. P2Y6 receptor inhibition aggravates ischemic brain injury by reducing microglial phagocytosis

    doi: 10.1111/cns.13296

    Figure Lengend Snippet: P2Y6 receptor antagonist MRS2578 inhibit MLCK expression in mice at 3 d after tMCAO and cultured microglia. A, (a) Representative fluorescence images of MLCK in Iba‐1 + cultured primary microglia. Scale bar = 25 μm. (b) Quantitative analysis of MLCK in each Iba1 + microglia. n = 4 per group. B, (a) Western blot for the expression of MLCK in the control, LPS+UDP and LPD+UDP+MRS2578 group. (b) Quantification for the intensity ratios of MLCK/β‐actin in the control, LPS+UDP and LPD+UDP+MRS2578 group. C, (a) Western blot for the expression of MLCK in the sham, saline, and MRS2578 group at 3 d after tMCAO. (b) Quantification for the intensity ratios of MLCK/β‐actin in the sham, saline, and MRS2578 group. Data were presented as mean ± SEM. n = 3‐4 per group. * P

    Article Snippet: Then, the sections were incubated with primary antibodies anti‐P2Y6 receptor (1:50, Alomone), Iba1 (1:200, Novusbio), GFAP (1:200, Millipore), Tuj‐1 (1:200, Millipore), NeuN (1:200, Millipore), or Lamp‐2 (1:200, Millipore) overnight at 4°C.

    Techniques: Expressing, Mouse Assay, Cell Culture, Fluorescence, Western Blot

    Inhibition of P2Y6 receptor activity reduced the phagocytic function of microglia after tMCAO. A, Representative fluorescence images of Iba1 + microglia (red) and TUNEL + apoptotic cells (green) in the peri‐infarct region of brain slice in the sham (a), saline (b), and MRS2578 (c) group at 3 d after tMCAO. Scale bar = 50 μm. White arrows indicated Iba1 + cells engulfed TUNEL + cells in the saline and MRS2578 group. B, Semiquantitative analysis of Iba1 + microglial phagocytosis for TUNEL + cells. n = 4. C, Representative fluorescence images of Iba1 + microglia (green), lysosome marker LAMP‐2 (purple), and TUNEL + apoptotic cells (red) in the peri‐infarct region of brain slice after tMCAO. 3D scanning with depth indicated Iba1 + cells phagocytosed TUNEL + cells and transferred them into lysosomes. Scale bar = 5 μm. D, Representative fluorescence image of Iba1 + microglia (green), NeuN + neurons (purple), and TUNEL + apoptotic cells (red) in the peri‐infarct region of brain slice after tMCAO. Scale bar = 5 μm. 3D scanning with depth indicated Iba1 + cells engulfed TUNEL + neurons

    Journal: CNS Neuroscience & Therapeutics

    Article Title: P2Y6 receptor inhibition aggravates ischemic brain injury by reducing microglial phagocytosis, et al. P2Y6 receptor inhibition aggravates ischemic brain injury by reducing microglial phagocytosis

    doi: 10.1111/cns.13296

    Figure Lengend Snippet: Inhibition of P2Y6 receptor activity reduced the phagocytic function of microglia after tMCAO. A, Representative fluorescence images of Iba1 + microglia (red) and TUNEL + apoptotic cells (green) in the peri‐infarct region of brain slice in the sham (a), saline (b), and MRS2578 (c) group at 3 d after tMCAO. Scale bar = 50 μm. White arrows indicated Iba1 + cells engulfed TUNEL + cells in the saline and MRS2578 group. B, Semiquantitative analysis of Iba1 + microglial phagocytosis for TUNEL + cells. n = 4. C, Representative fluorescence images of Iba1 + microglia (green), lysosome marker LAMP‐2 (purple), and TUNEL + apoptotic cells (red) in the peri‐infarct region of brain slice after tMCAO. 3D scanning with depth indicated Iba1 + cells phagocytosed TUNEL + cells and transferred them into lysosomes. Scale bar = 5 μm. D, Representative fluorescence image of Iba1 + microglia (green), NeuN + neurons (purple), and TUNEL + apoptotic cells (red) in the peri‐infarct region of brain slice after tMCAO. Scale bar = 5 μm. 3D scanning with depth indicated Iba1 + cells engulfed TUNEL + neurons

    Article Snippet: Then, the sections were incubated with primary antibodies anti‐P2Y6 receptor (1:50, Alomone), Iba1 (1:200, Novusbio), GFAP (1:200, Millipore), Tuj‐1 (1:200, Millipore), NeuN (1:200, Millipore), or Lamp‐2 (1:200, Millipore) overnight at 4°C.

    Techniques: Inhibition, Activity Assay, Fluorescence, TUNEL Assay, Slice Preparation, Marker

    P2Y6 receptor‐specific inhibitor MRS2578 blocked the phagocytosis of primary microglia under LPS UDP stimulation. A, Representative images of fluorescent latex beads (yellow) phagocytized by primary microglia under 12 h LPS (200 ng/mL) UDP (100 μmol/L) stimulation in the control, LPS+UDP, 1 μmol/L MRS, 2 μmol/L MRS, and 5 μmol/L MRS group. (a) Control group, only cultured by basic culture medium of microglia, (b) MRS2578‐free group, 200 ng/mL LPS, and 100 μmol/L UDP in basic culture medium of microglia, (c) 1 μmol/L MRS2578 group, 200 ng/mL LPS, 100 μmol/L UDP, and 1 μmol/L MRS2578 in basic culture medium of microglia, (d) 2 μmol/L MRS2578 group, 200 ng/mL LPS, 100 μmol/L UDP, and 2 μmol/L MRS2578 in basic culture medium of microglia, (e) 5 μmol/L MRS2578 group, 200 ng/mL LPS, 100 μmol/L UDP, and 5 μmol/L MRS2578 in basic culture medium of microglia. Scale bar = 10 μm. B, Quantification for the number of beads per microglia under LPS (200 ng/mL) UDP (100 μmol/L) stimulation (n = 3 per group). C, CCK‐8 assay in primary microglia without or with different concentrations MRS2578 under LPS+UDP stimulation for 12 h. D, (a) Representative flow cytometry analysis of microglia phagocytosis. Phagocytosis of fluorescein isothiocyanate (FITC)‐labeled latex beads by CD206 + and CD206 ‐ microglia. (Green, untreated control; Red, LPS+UDP treatment; Blue: LPS+UDP+1 μmol/L MRS2578 treatment); (b) Percentage of FITC + cells in CX 3 CR 1 + cells for total, CD206+, and CD206‐ microglia. E, (a) Representative diagrams of CD206 + /CX 3 CR 1 + cells in the control, LPS+UDP, LPS+UDP+MRS2578 group, (b) Percentage of CD206 + /CX 3 CR 1 + in the control, LPS+UDP, LPS+UDP+MRS2578 group. Data were presented as mean ± SEM. n = 3 per group. * P

    Journal: CNS Neuroscience & Therapeutics

    Article Title: P2Y6 receptor inhibition aggravates ischemic brain injury by reducing microglial phagocytosis, et al. P2Y6 receptor inhibition aggravates ischemic brain injury by reducing microglial phagocytosis

    doi: 10.1111/cns.13296

    Figure Lengend Snippet: P2Y6 receptor‐specific inhibitor MRS2578 blocked the phagocytosis of primary microglia under LPS UDP stimulation. A, Representative images of fluorescent latex beads (yellow) phagocytized by primary microglia under 12 h LPS (200 ng/mL) UDP (100 μmol/L) stimulation in the control, LPS+UDP, 1 μmol/L MRS, 2 μmol/L MRS, and 5 μmol/L MRS group. (a) Control group, only cultured by basic culture medium of microglia, (b) MRS2578‐free group, 200 ng/mL LPS, and 100 μmol/L UDP in basic culture medium of microglia, (c) 1 μmol/L MRS2578 group, 200 ng/mL LPS, 100 μmol/L UDP, and 1 μmol/L MRS2578 in basic culture medium of microglia, (d) 2 μmol/L MRS2578 group, 200 ng/mL LPS, 100 μmol/L UDP, and 2 μmol/L MRS2578 in basic culture medium of microglia, (e) 5 μmol/L MRS2578 group, 200 ng/mL LPS, 100 μmol/L UDP, and 5 μmol/L MRS2578 in basic culture medium of microglia. Scale bar = 10 μm. B, Quantification for the number of beads per microglia under LPS (200 ng/mL) UDP (100 μmol/L) stimulation (n = 3 per group). C, CCK‐8 assay in primary microglia without or with different concentrations MRS2578 under LPS+UDP stimulation for 12 h. D, (a) Representative flow cytometry analysis of microglia phagocytosis. Phagocytosis of fluorescein isothiocyanate (FITC)‐labeled latex beads by CD206 + and CD206 ‐ microglia. (Green, untreated control; Red, LPS+UDP treatment; Blue: LPS+UDP+1 μmol/L MRS2578 treatment); (b) Percentage of FITC + cells in CX 3 CR 1 + cells for total, CD206+, and CD206‐ microglia. E, (a) Representative diagrams of CD206 + /CX 3 CR 1 + cells in the control, LPS+UDP, LPS+UDP+MRS2578 group, (b) Percentage of CD206 + /CX 3 CR 1 + in the control, LPS+UDP, LPS+UDP+MRS2578 group. Data were presented as mean ± SEM. n = 3 per group. * P

    Article Snippet: Then, the sections were incubated with primary antibodies anti‐P2Y6 receptor (1:50, Alomone), Iba1 (1:200, Novusbio), GFAP (1:200, Millipore), Tuj‐1 (1:200, Millipore), NeuN (1:200, Millipore), or Lamp‐2 (1:200, Millipore) overnight at 4°C.

    Techniques: Cell Culture, CCK-8 Assay, Flow Cytometry, Labeling

    Inhibition of P2Y6 receptor activity exacerbated neurological function deficit and brain injury after tMCAO in mice. A, (a) Representative images of cresyl violet staining for brain infarct volume in the sham, saline, and MRS2578 group at 3 d after tMCAO. (b) Quantification of brain infarct volume in the sham, saline, and MRS2578 group at 3 d after tMCAO. (c) Quantification of the percentage of ipsilateral hemisphere volume/contralateral hemisphere volume in the sham, saline, and MRS2578 group at 3 d after tMCAO. B, (d) Representative images of cresyl violet staining for brain atrophy volume in the sham, saline, and MRS2578 group at 14 d after tMCAO. (e) Quantification of brain atrophy volume in the sham, saline, and MRS2578 group at 14 d after tMCAO. (f) Quantification of the percentage of ipsilateral hemisphere volume/contralateral hemisphere volume in the sham, saline, and MRS2578 group at 14 d after tMCAO. C. mNSS in the saline and MRS2578 group at 1, 3, 7, and 14 d after tMCAO. mNSS = modified neurological severity scores. D, Grid walking test in the saline and MRS2578 group at 3, 7, and 14 d after tMCAO. E, Body weight in the saline and MRS2578 group at 1, 3, 7, and 14 d after tMCAO. F, Survival curves of the saline and MRS2578 group within 3 d after tMCAO. Data were presented as mean ± SEM. n = 6‐11 per group. * P

    Journal: CNS Neuroscience & Therapeutics

    Article Title: P2Y6 receptor inhibition aggravates ischemic brain injury by reducing microglial phagocytosis, et al. P2Y6 receptor inhibition aggravates ischemic brain injury by reducing microglial phagocytosis

    doi: 10.1111/cns.13296

    Figure Lengend Snippet: Inhibition of P2Y6 receptor activity exacerbated neurological function deficit and brain injury after tMCAO in mice. A, (a) Representative images of cresyl violet staining for brain infarct volume in the sham, saline, and MRS2578 group at 3 d after tMCAO. (b) Quantification of brain infarct volume in the sham, saline, and MRS2578 group at 3 d after tMCAO. (c) Quantification of the percentage of ipsilateral hemisphere volume/contralateral hemisphere volume in the sham, saline, and MRS2578 group at 3 d after tMCAO. B, (d) Representative images of cresyl violet staining for brain atrophy volume in the sham, saline, and MRS2578 group at 14 d after tMCAO. (e) Quantification of brain atrophy volume in the sham, saline, and MRS2578 group at 14 d after tMCAO. (f) Quantification of the percentage of ipsilateral hemisphere volume/contralateral hemisphere volume in the sham, saline, and MRS2578 group at 14 d after tMCAO. C. mNSS in the saline and MRS2578 group at 1, 3, 7, and 14 d after tMCAO. mNSS = modified neurological severity scores. D, Grid walking test in the saline and MRS2578 group at 3, 7, and 14 d after tMCAO. E, Body weight in the saline and MRS2578 group at 1, 3, 7, and 14 d after tMCAO. F, Survival curves of the saline and MRS2578 group within 3 d after tMCAO. Data were presented as mean ± SEM. n = 6‐11 per group. * P

    Article Snippet: Then, the sections were incubated with primary antibodies anti‐P2Y6 receptor (1:50, Alomone), Iba1 (1:200, Novusbio), GFAP (1:200, Millipore), Tuj‐1 (1:200, Millipore), NeuN (1:200, Millipore), or Lamp‐2 (1:200, Millipore) overnight at 4°C.

    Techniques: Inhibition, Activity Assay, Mouse Assay, Staining, Modification

    Inhibiting the P2Y6 receptor activity had no effect on the expression of inflammatory cytokines and neutrophil infiltration. A, Quantification of RT‐PCR analysis for the expression of IL‐1α (a), IL‐1β (b), IL‐6 (c), IL‐10 (d), TNF‐α (e), and TGF‐β (f) at 3 d after tMCAO in the sham, saline, and MRS2578 group. The data in sham group were normalized to 1. B, Western blot for the expression of MPO in the brain of sham, saline, and MRS2578 groups at 3 d after tMCAO (g). Quantification of the intensity ratios of MPO/ β‐actin (h) in the sham, saline, and MRS2578 group. The data in the saline group were normalized to 1. C, Quantification of RT‐PCR analysis for the expression of IL‐1α (a), IL‐1β (b), IL‐6 (c), IL‐10(d), TNF‐α (e), and TGF‐β (f) after 1 h OGD and 11 h reoxygenation in the control, OGD and OGD+MRS2578 group. The data in the control group were normalized to 1. Data were presented as mean ± SEM. n = 3‐4 per group. * P

    Journal: CNS Neuroscience & Therapeutics

    Article Title: P2Y6 receptor inhibition aggravates ischemic brain injury by reducing microglial phagocytosis, et al. P2Y6 receptor inhibition aggravates ischemic brain injury by reducing microglial phagocytosis

    doi: 10.1111/cns.13296

    Figure Lengend Snippet: Inhibiting the P2Y6 receptor activity had no effect on the expression of inflammatory cytokines and neutrophil infiltration. A, Quantification of RT‐PCR analysis for the expression of IL‐1α (a), IL‐1β (b), IL‐6 (c), IL‐10 (d), TNF‐α (e), and TGF‐β (f) at 3 d after tMCAO in the sham, saline, and MRS2578 group. The data in sham group were normalized to 1. B, Western blot for the expression of MPO in the brain of sham, saline, and MRS2578 groups at 3 d after tMCAO (g). Quantification of the intensity ratios of MPO/ β‐actin (h) in the sham, saline, and MRS2578 group. The data in the saline group were normalized to 1. C, Quantification of RT‐PCR analysis for the expression of IL‐1α (a), IL‐1β (b), IL‐6 (c), IL‐10(d), TNF‐α (e), and TGF‐β (f) after 1 h OGD and 11 h reoxygenation in the control, OGD and OGD+MRS2578 group. The data in the control group were normalized to 1. Data were presented as mean ± SEM. n = 3‐4 per group. * P

    Article Snippet: Then, the sections were incubated with primary antibodies anti‐P2Y6 receptor (1:50, Alomone), Iba1 (1:200, Novusbio), GFAP (1:200, Millipore), Tuj‐1 (1:200, Millipore), NeuN (1:200, Millipore), or Lamp‐2 (1:200, Millipore) overnight at 4°C.

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

    RT-PCR and sequence analysis of porcine P2Y2 , P2Y4 and P2Y6 receptors

    Journal:

    Article Title: Evidence for the Expression of Multiple Uracil Nucleotide-Stimulated P2 Receptors Coupled to Smooth Muscle Contraction in Porcine Isolated Arteries

    doi: 10.1038/sj.bjp.0707120

    Figure Lengend Snippet: RT-PCR and sequence analysis of porcine P2Y2 , P2Y4 and P2Y6 receptors

    Article Snippet: Antibodies directed against sequence-specific peptides from P2Y2 , P2Y4 and P2Y6 receptors were from Alomone Labs/Caltag-Medsystems (Buckingham, UK), while HRP-conjugated goat anti-rabbit antibody was from DAKO/Cytomation (Ely, UK).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Sequencing

    RT-PCR and immunoblot analysis of P2Y receptor expression in pig coronary and ear arteries. Agarose gel electrophoresis ( a–c ) showing size marker (MW) and PCR products from coronary (PCA) and ear (PEA) arteries or no template (CON) for p2ry2 (

    Journal:

    Article Title: Evidence for the Expression of Multiple Uracil Nucleotide-Stimulated P2 Receptors Coupled to Smooth Muscle Contraction in Porcine Isolated Arteries

    doi: 10.1038/sj.bjp.0707120

    Figure Lengend Snippet: RT-PCR and immunoblot analysis of P2Y receptor expression in pig coronary and ear arteries. Agarose gel electrophoresis ( a–c ) showing size marker (MW) and PCR products from coronary (PCA) and ear (PEA) arteries or no template (CON) for p2ry2 (

    Article Snippet: Antibodies directed against sequence-specific peptides from P2Y2 , P2Y4 and P2Y6 receptors were from Alomone Labs/Caltag-Medsystems (Buckingham, UK), while HRP-conjugated goat anti-rabbit antibody was from DAKO/Cytomation (Ely, UK).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Agarose Gel Electrophoresis, Marker, Polymerase Chain Reaction

    Subcellular distribution of P2Y2, P2Y4, and P2Y6 receptors in MEFs. WT or KI MEFs were cultured and, after changing the medium, were maintained in the absence or presence of 1 μ M DFU for 2 h. Cells were fixed with paraformaldehyde (4%; pH 7.2) and permeabilized with cold methanol at RT. After incubation with anti-P2Y2, anti-P2Y4 and anti-P2Y6 antibodies (1 : 500) overnight at 4°C, cells were visualized by confocal microscopy using a FITC-conjugated secondary Ab (Alexa-Fluor 488, 1 : 1000). Nuclei were stained with Hoechst 33258. Coverslips were mounted in Prolong Gold antifade reagent (Molecular Probes) and the intensity of the fluorescence was measured using Image J software (NIH, Bethesda, MD, USA). Results show the mean + SD of three experiments. ** P

    Journal: Mediators of Inflammation

    Article Title: Sustained Release of Prostaglandin E2 in Fibroblasts Expressing Ectopically Cyclooxygenase 2 Impairs P2Y-Dependent Ca2+-Mobilization

    doi: 10.1155/2014/832103

    Figure Lengend Snippet: Subcellular distribution of P2Y2, P2Y4, and P2Y6 receptors in MEFs. WT or KI MEFs were cultured and, after changing the medium, were maintained in the absence or presence of 1 μ M DFU for 2 h. Cells were fixed with paraformaldehyde (4%; pH 7.2) and permeabilized with cold methanol at RT. After incubation with anti-P2Y2, anti-P2Y4 and anti-P2Y6 antibodies (1 : 500) overnight at 4°C, cells were visualized by confocal microscopy using a FITC-conjugated secondary Ab (Alexa-Fluor 488, 1 : 1000). Nuclei were stained with Hoechst 33258. Coverslips were mounted in Prolong Gold antifade reagent (Molecular Probes) and the intensity of the fluorescence was measured using Image J software (NIH, Bethesda, MD, USA). Results show the mean + SD of three experiments. ** P

    Article Snippet: Antibodies against P2Y2, P2Y4, and P2Y6 receptors were from Alomone Labs (Jerusalem, Israel) and other antibodies were from Santa Cruz Biotech (Santa Cruz, CA, USA), from Cell Signaling (Danvers, MA, USA), or from the sources previously described [ ].

    Techniques: Cell Culture, Incubation, Confocal Microscopy, Staining, Fluorescence, Software

    Characterization of EP1–4 and P2Y2–P2Y4 expression and effect of ionophores on Ca 2+ -mobilization in MEF cells. The expression levels of the prostaglandin receptors EP1–4 and the levels of P2Y2 and P2Y4 were determined by qPCR (a-b). The response to 1 μ M ionomycin (c) and 500 nM thapsigargin (d) on Ca 2+ -mobilization was determined in MEFs overexpressing COX-2, using the dual excitation 340/380 nm protocol as described in Section 2 . MEFs KI were washed with fresh medium to remove PGE 2 accumulated and maintained in the absence or presence of 1 μ M DFU and 5 μ M PGE 2 . Different extracellular concentrations of calcium were used. Results show the mean + SD of three experiments (a-b) or a representative trace (c-d). * P

    Journal: Mediators of Inflammation

    Article Title: Sustained Release of Prostaglandin E2 in Fibroblasts Expressing Ectopically Cyclooxygenase 2 Impairs P2Y-Dependent Ca2+-Mobilization

    doi: 10.1155/2014/832103

    Figure Lengend Snippet: Characterization of EP1–4 and P2Y2–P2Y4 expression and effect of ionophores on Ca 2+ -mobilization in MEF cells. The expression levels of the prostaglandin receptors EP1–4 and the levels of P2Y2 and P2Y4 were determined by qPCR (a-b). The response to 1 μ M ionomycin (c) and 500 nM thapsigargin (d) on Ca 2+ -mobilization was determined in MEFs overexpressing COX-2, using the dual excitation 340/380 nm protocol as described in Section 2 . MEFs KI were washed with fresh medium to remove PGE 2 accumulated and maintained in the absence or presence of 1 μ M DFU and 5 μ M PGE 2 . Different extracellular concentrations of calcium were used. Results show the mean + SD of three experiments (a-b) or a representative trace (c-d). * P

    Article Snippet: Antibodies against P2Y2, P2Y4, and P2Y6 receptors were from Alomone Labs (Jerusalem, Israel) and other antibodies were from Santa Cruz Biotech (Santa Cruz, CA, USA), from Cell Signaling (Danvers, MA, USA), or from the sources previously described [ ].

    Techniques: Expressing, Real-time Polymerase Chain Reaction