rabbit anti ca v β3 cacnb3  (Alomone Labs)

Journal: Nature Communications

Article Title: CYFIP1 governs the development of cortical axons by modulating calcium availability

doi: 10.1038/s41467-025-65801-0

Figure Lengend Snippet: a CYFIP1 RNA immunoprecipitation (RNA-IP) from DIV 3 WT cortical neurons. Histogram showing relative enrichment of the mRNAs over the non-specific IgG, measured by RT-qPCR of the eluate. The values were normalized for the input and mHprt1 mRNA and expressed as fold change over the non-specific IgG of each mRNA ( n = 4 embryos; mean ± SEM; One-Way ANOVA p < 0.0001; mMap1b mRNA p = 0.0390, mCacna1c mRNA p = 0.0054, mCacna1e mRNA p = 0.0078, mCacna1i mRNA p = 0.0009, mCacng2 mRNA p = 0.9997, mCacnb3 mRNA p = 0.9983). b Total mRNA levels of the Ca 2+ channels in DIV 3 WT and Cyfip1 +/- cortical neurons. Histograms represent mCacna1c , mCacna1e , mCacna1i , mCacng2, mCacnb3 and mCyfip1 mRNA levels, normalized to mH3f3 levels and expressed as a fold change over WT (WT n = 6/7 embryos, Cyfip1 +/- n = 7 embryos; mean ± SEM; Two-tailed Multiple Mann-Whitney test, mCacna1c mRNA p = 0.0766, mCacna1e mRNA p = 0.0435, mCacna1i mRNA p = 0.0202, mCacng2 mRNA p = 0.6282, mCacnb3 mRNA p = 0.5343, mCyfip1 mRNA p = 0.0034). c Left, representative Western Blot showing CYFIP1, Ca V 1.2 (CACNA1C), Ca V 2.3 (CACNA1E), Ca V 3.3 (CACNA1I), Ca V γ2 (CACNG2/Stargazin) and Ca V β3 (CACNB3) in membrane-enriched fractions from WT and Cyfip1 +/- DIV 3 cortical neurons. The molecular weight of each protein is indicated in kDa. Right, histogram representing Ca V 1.2, Ca V 2.3, Ca V 3.3, Ca V γ2, Ca V β3 and CYFIP1 protein expression levels in membrane-enriched fractions from WT and Cyfip1 +/- DIV 3 cortical neurons. Protein levels were normalized to Coomassie staining (WT n = 4 embryos, Cyfip1 +/- n = 7/8 embryos; mean ± SEM; Two-tailed Multiple unpaired t -test, Ca V 1.2 p = 0.0338, Ca V 2.3 p = 0.0281, Ca V 3.3 p = 0.0129, Ca V γ2 p = 0.2574, Ca V β3 p = 0.6259, CYFIP1 p = 0.0137). d–f Representative images from WT and Cyfip1 +/- DIV 3 cortical neurons stained for Ca V 1.2, Ca V 2.3, Ca V 3.3 (magenta) and βIII-Tubulin (green) (scale bar 20 μm). Histograms show the fluorescence intensity of each calcium channel normalized to βIII-Tubulin in the total neuron (left) and in the axon (right), expressed as a percentage over WT (Ca V 1.2: WT n = 4 embryos, Cyfip1 +/- n = 5 embryos; mean ± SEM; Two-tailed Mann-Whitney test, total p = 0.1111, axon p = 0.4127; Ca V 2.3: WT n = 5 embryos, Cyfip1 +/- n = 4 embryos; mean ± SEM; Two-tailed Mann-Whitney test, total p = 0.0635, axon p = 0.0159; Ca V 3.3: WT n = 4 embryos, Cyfip1 +/- n = 4 embryos; mean ± SEM; Two-tailed Mann-Whitney test, total p = 0.0286, axon p = 0.0286). Source data are provided as a Source Data file.

Article Snippet: Membranes were incubated with the following antibodies rabbit anti-CYFIP1 (1:1000; Sigma-Aldrich, #AB6046), rabbit anti-Ca V 1.2 (CACNA1C) (1:500, Alomone Labs, #ACC003), rabbit anti-Ca V 2.3 (CACNA1E) (1:500, Alomone Labs, #ACC006), rabbit anti-Ca V 3.3 (CACNA1I) (1:500, Alomone Labs, #ACC009), rabbit anti-Stargazin (Ca V γ2/CACNG2) (1:500, Alomone Labs, #ACC012) and rabbit anti-Ca V β3 (CACNB3) (1:500, Alomone Labs, #ACC008), HuD/Elavl4 (1:500, Santa Cruz, #sc-48421) and HuR/Elavl1 (1:500, Santa Cruz, #sc-5261).

Techniques: RNA Immunoprecipitation, Quantitative RT-PCR, Two Tailed Test, MANN-WHITNEY, Western Blot, Membrane, Molecular Weight, Expressing, Staining, Fluorescence

Journal: Cells

Article Title: Sympathetic Stimulation Upregulates the Ca 2+ Channel Subunit, Ca V α2δ1, via the β1 and ERK 1/2 Pathway in Neonatal Ventricular Cardiomyocytes

doi: 10.3390/cells11020188

Figure Lengend Snippet: Expression of Ca V α2δ1 ( A ), Ca V α1C ( C ), Ca V β3 ( E ), and Ca V β2 ( G ) in neonatal rat cardiomyocytes. Total proteins were extracted from neonatal rat ventricular cardiomyocytes. Proteins were separated on an 8% SDS-polyacrylamide gel, transferred to a nitrocellulose membrane, and probed with anti-Ca V α2δ1, anti-Ca V α1C, anti-Ca V β3, anti-Ca V β2, and anti-GAPDH antibodies overnight and then incubated with HRP-conjugated goat anti-rabbit secondary antibody. Lanes were loaded with 30 μg of proteins. NE: cardiomyocytes treated with 1 µM norepinephrine. Graph showing the total protein expression of Ca V α2δ1 ( B ), Ca V α1C ( D ), Ca V β3 ( F ), and Ca V β2 ( H ) normalized to GAPDH. * p < 0.01 vs. Basal. Statistical analysis was performed using one-way ANOVA.

Article Snippet: Antibodies used were rabbit anti-Ca V α2δ1 (1:1000, Alomone ACC-015), rabbit Ca V α1C (1:1000, Alomone ACC-003), rabbit Ca V β2 (1:5000, Alomone ACC-105), rabbit Ca V β3 (1:500, Alomone ACC-008), rabbit Phospho-ERK 1/2 (Cell Signaling Technology, Danvers, MA, USA, 9101S), Total ERK 1/2 (Cell Signaling Technology, 137F5) and rabbit GAPDH (Jackson, West Grove, PA, USA, 111-035-144).

Techniques: Expressing, Incubation

Journal: Cells

Article Title: Sympathetic Stimulation Upregulates the Ca 2+ Channel Subunit, Ca V α2δ1, via the β1 and ERK 1/2 Pathway in Neonatal Ventricular Cardiomyocytes

doi: 10.3390/cells11020188

Figure Lengend Snippet: Expression of Ca V α2δ1 ( A ) and Ca V β3 ( C ) in neonatal rat cardiomyocytes. Total proteins were extracted from neonatal rat ventricular cardiomyocytes. Cells were treated with 1 µM Norepinephrine (NE) for 24 h or pre-treated with metoprolol tartrate (β1-blocker) for 1 h followed by treatment with NE for 24 h. Proteins were separated on an 8% SDS-polyacrylamide gel, transferred to a nitrocellulose membrane, and probed with anti-Ca V α2δ1, anti-Ca V β3, and anti-GAPDH antibodies overnight and then incubated with HRP-conjugated goat anti-rabbit secondary antibody. Lanes were loaded with 30 μg of proteins. NE: cardiomyocytes treated with 1 µM norepinephrine. 24 h + metoprolol tartrate: Cardiomyocytes pre-treated with metoprolol tartrate (β1-blocker) for 1 h, followed by treatment with NE for 24 h. ( B ) Graph showing the total protein expression of Ca V α2δ1 normalized to GAPDH. * p < 0.01 vs. Basal untreated NRVMs; † p <0.01 vs. NE 24 h. (4 cardio-preparations). (D) Graph showing the total protein expression of Ca V β3 normalized to GAPDH. * p < 0.01 vs. Basal untreated NRVMs (4 cardio-preparations). Statistical analysis was performed using one-way ANOVA.

Article Snippet: Antibodies used were rabbit anti-Ca V α2δ1 (1:1000, Alomone ACC-015), rabbit Ca V α1C (1:1000, Alomone ACC-003), rabbit Ca V β2 (1:5000, Alomone ACC-105), rabbit Ca V β3 (1:500, Alomone ACC-008), rabbit Phospho-ERK 1/2 (Cell Signaling Technology, Danvers, MA, USA, 9101S), Total ERK 1/2 (Cell Signaling Technology, 137F5) and rabbit GAPDH (Jackson, West Grove, PA, USA, 111-035-144).

Techniques: Expressing, Incubation