p2y2  (Alomone Labs)

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

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

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

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

Journal: Purinergic Signalling

Article Title: Impact of CD39 and purinergic signalling on the growth and metastasis of colorectal cancer

doi: 10.1007/s11302-011-9228-9

Figure Lengend Snippet: Purinergic studies in human CRC. Protein expression levels of CD39, P2X7 and P2Y2 ( a – c ) and localization of P2Y2 ( d , e ) was determined in human CRC. Changes in protein levels were observed for P2Y2 ( c ), which was significantly less in T3N + M0

Article Snippet: To validate P2 receptor protein expression (P2X7, P2Y2), primary antibodies against P2X7 and P2Y2 were blocked with the specific antigen, as provided by the manufacturer (Alomone labs).

Techniques: Expressing

Journal: Purinergic Signalling

Article Title: Impact of CD39 and purinergic signalling on the growth and metastasis of colorectal cancer

doi: 10.1007/s11302-011-9228-9

Figure Lengend Snippet: CD39 and P2 receptors in CRC. a In orthotopic tumors, CD39, P2X7 and P2Y2 mRNA appeared at comparable levels to those seen in control livers. b As expected, mouse CD39 mRNA was significantly lower expressed in genetically deficient over wild-type and

Article Snippet: To validate P2 receptor protein expression (P2X7, P2Y2), primary antibodies against P2X7 and P2Y2 were blocked with the specific antigen, as provided by the manufacturer (Alomone labs).

Techniques:

Journal: Purinergic Signalling

Article Title: Bone turnover is altered in transgenic rats overexpressing the P2Y2 purinergic receptor

doi: 10.1007/s11302-017-9582-3

Figure Lengend Snippet: a Fold change in expression of P2Y2 mRNA in brain and lung tissue ( n = 3) as well as calvarial osteoblasts ( n = 4) of P2Y2R-Tg rats relative to Wt ( n = 3 (brain and lung); n = 6 (calvarial osteoblasts)). b P2Y2R protein expression in calvarial osteoblasts from WT ( n = 4) and P2Y2R-Tg ( n = 4) rats as measured by Western blotting. All bands are normalized to histone H3 expression, and mean expression in Wt cells is set at 100%. Lower panel shows membrane images of each individual experiment. Error bars represent SEM. TG P2Y2R transgenic, Wt wild-type

Article Snippet: They were incubated overnight using rabbit anti-P2Y2 (APR-010, Alomone Labs, 1:1000) or monoclonal rabbit anti-histone H3 (#4499S, Cell Signaling, 1:1000) in blocking buffer at 4 °C on a rotor; briefly washed twice in TBS-T and then incubated with HRP-conjugated donkey anti-rabbit IgG antibody (#NA934V, GE Healthcare, 1:40.000 in blocking buffer) at RT on a rotor; washed in large volumes of TBS-T; and finally developed using ECL Select Western Blotting Detection Kit (GE Healthcare) for 5 min at RT.

Techniques: Expressing, Western Blot, Transgenic Assay