P2X2 (P2RX2) Guinea Pig Polyclonal Antibody

CAT#: GP14106-150

P2X2 (P2RX2) guinea pig polyclonal antibody

Product Images

P2x2 staining of Cerebellar Purjinke cell layer. ~50 μm sections from 3 brains from P15 rats. http://stke.sciencemag.org/cgi/data/sigtrans;1/41/ra8/DC1/1

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  P2x2 staining of Cerebellar Purjinke cell layer. ~50 μm sections from 3 brains from P15 rats. http://stke.sciencemag.org/cgi/data/sigtrans;1/41/ra8/DC1/1

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  The constitutive interaction between P2X2 receptors and VILIP1 is calcium independent. Calcium dependence of the interaction between P2X2 receptors and VILIP1 was assessed by coimmunoprecipitation from HEK cells expressing P2X2-FLAG receptors and either VILIP1or VILIP1 EF-hand mutants. The effect of calcium on the interaction was tested before cell lysis by increasing intracellular calcium with ATP application or ionomycin treatment, and also during the immunoprecipitation steps. In the latter case, experiments were carried in the absence of calcium and in the presence of 5 mM EDTA or in the absence of EDTA and in the presence of 500μM calcium. P2X2 receptors were immunoprecipitated using a FLAG antibody conjugated to agarose beads. Bound proteins were eluted by competition using a FLAG peptide. As shown in the top panel, no difference in P2X2-FLAG-VILIP1 interaction was observed in the presence or the absence of calcium in the immunoprecipitation buffer. In addition, stimulation of transfected cells with ATP (100μM, 5 minutes) or with ionomycin (2μM, 5 minutes) prior to lysis did not affect the interaction between the two proteins. A similar observation was made when the VILIP1 EF hand mutant was co-immunoprecipitated. No immunoprecipitation was observed when VILIP1 was transfected alone. The bottom panel shows the total protein input used for immunoprecipitation. www.sciencesignaling.org/cgi/content/full/1/41/ra8/DC1

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  The constitutive interaction between P2X2 receptors and VILIP1 is calcium independent. Calcium dependence of the interaction between P2X2 receptors and VILIP1 was assessed by coimmunoprecipitation from HEK cells expressing P2X2-FLAG receptors and either VILIP1or VILIP1 EF-hand mutants. The effect of calcium on the interaction was tested before cell lysis by increasing intracellular calcium with ATP application or ionomycin treatment, and also during the immunoprecipitation steps. In the latter case, experiments were carried in the absence of calcium and in the presence of 5 mM EDTA or in the absence of EDTA and in the presence of 500μM calcium. P2X2 receptors were immunoprecipitated using a FLAG antibody conjugated to agarose beads. Bound proteins were eluted by competition using a FLAG peptide. As shown in the top panel, no difference in P2X2-FLAG-VILIP1 interaction was observed in the presence or the absence of calcium in the immunoprecipitation buffer. In addition, stimulation of transfected cells with ATP (100μM, 5 minutes) or with ionomycin (2μM, 5 minutes) prior to lysis did not affect the interaction between the two proteins. A similar observation was made when the VILIP1 EF hand mutant was co-immunoprecipitated. No immunoprecipitation was observed when VILIP1 was transfected alone. The bottom panel shows the total protein input used for immunoprecipitation. www.sciencesignaling.org/cgi/content/full/1/41/ra8/DC1

Specifications

Product Data

• Applications: IF, IHC, WB
• Reactivities: Human, Monkey, Rat
• Host: Guinea Pig
• Clonality: Polyclonal
• Formulation: State: Serum
• Conjugation: Unconjugated
• Gene Name: Homo sapiens purinergic receptor P2X 2 (P2RX2), transcript variant 6
• Database Link:
  Entrez Gene 114115 RatEntrez Gene 22953 Human
  UniProt ID Q9UBL9
• Synonyms: P2X purinoceptor 2, ATP receptor, Purinergic receptor

Reference Data

• Protein Families: Druggable Genome, Ion Channels: ATP Receptors, Transmembrane
• Protein Pathways: Calcium signaling pathway, Neuroactive ligand-receptor interaction