rabbit polyclonal anti stargazin γ2  (Alomone Labs)

Journal: Scientific Reports

Article Title: Multiple H+ sensors mediate the extracellular acidification-induced [Ca2+]i elevation in cultured rat ventricular cardiomyocytes

doi: 10.1038/srep44951

Figure Lengend Snippet: Schematic diagram of mechanisms for the extracellular acidification inducing “Ca 2+ transients” in cultured rat cardiac myocytes. There are two components for Ca 2+ elevation in response to elevated external protons, one is ASIC/TRPV1 channel and another is OGR1protein. On one side, ASICs and TRPV1 can be activated by acidosis solution and mediate extracellular Ca 2+ entry. On the other side, OGR1 mediates mobilization of intracellular Ca 2+ from SR via OGR1-PLC-IP3-IP3R signaling pathway.

Article Snippet: After blocking, the proteins were probed with the appropriate primary antibodies against ASIC1, ASIC2a, ASIC3, TRPV1 (Alomone labs, Jerusalem, Israel.ASC-014, ASC-012, ASC-018, ACC-030, all in 1:200 dilution) and OGR1 (Santa Cruz Biotechnology, USA. sc-98437).

Techniques: Cell Culture, Planar Chromatography

Journal: Scientific Reports

Article Title: Multiple H+ sensors mediate the extracellular acidification-induced [Ca2+]i elevation in cultured rat ventricular cardiomyocytes

doi: 10.1038/srep44951

Figure Lengend Snippet: Functional expression of TRPV1 channel in rat cardiomyocytes. ( a ) RT-PCR detection of TRPV1 mRNA expressions in cultured rat ventricular cardiocytes. GAPDH were used as positive controls. M: marker; 1, 4: cardiomyocytes; 2: cortex; 3: negative control. ( b ) Western blotting indicating the protein expression of TRPV1 in cultured ventricular cardiocytes of rat. TRPV1 peptide was used as negative control. The blots were cropped from Supplementary Fig. S1b . ( c ) Double immunostaining of TRPV1 ( green ) and nucleus ( blue , marker: Hoechst33258) in rat cultured cardiomyocytes. NC: pretreatment with immunogenic peptide as negative control. Scale bars: 20 μm. Above all data were represented from three similar independent experiments. ( d ) TRPV1-like currents reversibly inhibited by CPZ (20 μM, n = 3 cells) in cultured and acute isolated rat cardiomyocytes. ( e ) Representative TRPV1 current traces evoked by indicated pH solutions from pH 7.4 in cultured rat cardiomyocytes. “—” indicates the duration of pH = 7.0, 6.0 or 5.0. ( f ) Representative [Ca 2+ ] i responses and quantitative analysis of normalized Fura-2/AM fluorescence induced by capsaicin (10 μM, n = 16 cells) and CPZ (20 μM) + capsaicin (10 μM, n = 16 cells). Data were shown as mean ± s.e.m. ** P

Article Snippet: After blocking, the proteins were probed with the appropriate primary antibodies against ASIC1, ASIC2a, ASIC3, TRPV1 (Alomone labs, Jerusalem, Israel.ASC-014, ASC-012, ASC-018, ACC-030, all in 1:200 dilution) and OGR1 (Santa Cruz Biotechnology, USA. sc-98437).

Techniques: Functional Assay, Expressing, Reverse Transcription Polymerase Chain Reaction, Cell Culture, Marker, Negative Control, Western Blot, Double Immunostaining, Isolation, Fluorescence

Journal: Scientific Reports

Article Title: Multiple H+ sensors mediate the extracellular acidification-induced [Ca2+]i elevation in cultured rat ventricular cardiomyocytes

doi: 10.1038/srep44951

Figure Lengend Snippet: Inhibitory effect of ASIC and TRPV1 inhibitors on extracellular acid solution-induced [Ca 2+ ] i elevation in cultured rat ventricular cardiomyocytes. ( a ) Representative [Ca 2+ ] i responses and quantitative analysis of normalized Fura-2/AM fluorescence induced by pH 6.0 (n = 15 cells) and pH 5.0 (n = 19 cells) solutions. Data were expressed as mean ± s.e.m (** P

Article Snippet: After blocking, the proteins were probed with the appropriate primary antibodies against ASIC1, ASIC2a, ASIC3, TRPV1 (Alomone labs, Jerusalem, Israel.ASC-014, ASC-012, ASC-018, ACC-030, all in 1:200 dilution) and OGR1 (Santa Cruz Biotechnology, USA. sc-98437).

Techniques: Cell Culture, Fluorescence