rabbit anti cav β3  (Alomone Labs)

Journal: bioRxiv

Article Title: CACNB4 overexpression decreases dendritic spine density in sex-specific manner

doi: 10.1101/2022.02.02.478824

Figure Lengend Snippet: β subunit protein levels. A) (Left) Mean optical density (a.u.) of β4 was significantly lower in P84, relative to P28 male and female mice (F=9.635, DF=3, p=0.013), with no significant sex or age by sex interactions. Error bars = SEM. (Right) Representative blot showing β4 bands at ~51-55kD. B) (Left) Mean optical density (a.u.) of β1 was significantly lower in P84, relative to P28 male and female mice (F=21.499, DF=3, p=0.001). There was also a main effect of sex (F=6.944, DF=3, p=0.027) and a significant age by sex interaction (F=6.835, DF=3, p=0 .028). Main effect of sex was significant at P28 (F=12.138, DF=1, p=0.040) but not P84 (F=0.016, DF=1, p=0.906). Error bars = SEM. (Right) Representative blot showing β1 bands at ~50-80kD. C) (Left) Neither age nor sex significantly impacted mean optical density (a.u.) of β3 and there was not a significant sex by age interaction. Error bars = SEM. (Right) Representative blot showing β3 bands at ~55kD (molecular weight of the predominant β3 isoform).

Article Snippet: The membrane was incubated overnight in Pierce SuperBlock Blocking Buffer and validated primary antibodies: either mouse anti-CaV β1 (Neuromab #73-052 1:100), rabbit anti-CaV β3 (Alomone Labs #AAC-008, 1:800) or mouse anti-CaVβ4 (Neuromab #75-054, 1:1000) and rabbit anti-β-tubulin (Abcam #ab6046, 1:600,000).

Techniques: Mouse Assay, Molecular Weight

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

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