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    Alomone Labs anti machr m3
    Expressions of mAChRs and key mediators within their downstream signaling pathway in Lop+TEE treated constipation rats. Expression levels of mAChRs and key mediators, including mAChR M2, mAChR M3, Gα, PKC, p-PKC, PI3K and p-PI3K, in the mAChR M2 and M3 signaling pathway were measured by Western blot analysis using specific primary antibodies and HRP-labeled anti-rabbit IgG antibody. After the intensity of each band was determined using an imaging densitometer, relative levels of the seven proteins were calculated based on the intensity of actin. Four to six rats per group were used in the preparation of the total tissue homogenate, and Western blot analyses were assayed in duplicate in each sample. The data are reported as the mean ± SD. *, p
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    Expressions of mAChRs and key mediators within their downstream signaling pathway in Lop+TEE treated constipation rats. Expression levels of mAChRs and key mediators, including mAChR M2, mAChR M3, Gα, PKC, p-PKC, PI3K and p-PI3K, in the mAChR M2 and M3 signaling pathway were measured by Western blot analysis using specific primary antibodies and HRP-labeled anti-rabbit IgG antibody. After the intensity of each band was determined using an imaging densitometer, relative levels of the seven proteins were calculated based on the intensity of actin. Four to six rats per group were used in the preparation of the total tissue homogenate, and Western blot analyses were assayed in duplicate in each sample. The data are reported as the mean ± SD. *, p

    Journal: PLoS ONE

    Article Title: Antioxidant activity and laxative effects of tannin-enriched extract of Ecklonia cava in loperamide-induced constipation of SD rats

    doi: 10.1371/journal.pone.0246363

    Figure Lengend Snippet: Expressions of mAChRs and key mediators within their downstream signaling pathway in Lop+TEE treated constipation rats. Expression levels of mAChRs and key mediators, including mAChR M2, mAChR M3, Gα, PKC, p-PKC, PI3K and p-PI3K, in the mAChR M2 and M3 signaling pathway were measured by Western blot analysis using specific primary antibodies and HRP-labeled anti-rabbit IgG antibody. After the intensity of each band was determined using an imaging densitometer, relative levels of the seven proteins were calculated based on the intensity of actin. Four to six rats per group were used in the preparation of the total tissue homogenate, and Western blot analyses were assayed in duplicate in each sample. The data are reported as the mean ± SD. *, p

    Article Snippet: Proteins (30 μg) were subjected to 4–20% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) for 3 h, and the resolved proteins were transferred to nitrocellulose membranes for 2 h at 40 V. The membranes were then probed with the following primary antibodies, overnight at 4°C: anti-Gα (ab128900, 1:1,000, Abcam, Cambridge, UK), anti-mAChR M2 (AMR-002, 1:1,000, Alomone Labs, Jerusalem, Israel), anti-mAChR M3 (AMR-006, 1:1,000, Alomone Labs), anti-PKC (2058s, 1:1,000, Cell Signaling Technology Inc., Danvers, MA, USA), anti-p-PKC (9376s, 1:1,000, Cell Signaling Technology Inc.), anti-PI-3K (4292s, 1:1,000, Cell Signaling Technology Inc.), anti-p-PI3K (4228s, 1:1,000, Cell Signaling Technology Inc.), anti-SOD (ab13498, 1:1,000, Abcam), anti-Nrf2 (ab137550, 1:1,000, Abcam), anti-p-Nrf2 (PA5-67520, 1:1,000, Invitrogen Co., Carlsvad, CA, USA), or anti-actin (4967s, 1:3,000, Sigma-Aldrich Co.).

    Techniques: Expressing, Western Blot, Labeling, Imaging

    Repression of P2rx1 by progressive vascular endothelial damage. ( a – c ) RQ-PCR ( a ) and western blot ( b , c ) analysis of the expressions of the cholinergic receptor muscarinic 2 ( Chrm2 ), cholinergic receptor muscarinic 3 ( Chrm3 ), and purinergic receptor P2X 1 ( P2rx1 ) genes in the bladders of rats in the indicated groups. Expression levels of the indicated transcripts are presented as % Gapdh . For western blot analysis, β-actin was used as a loading control, and the expression levels of the indicated proteins were normalized to the β-actin value. ( d , e ) Representative images ( d ) and quantification analysis ( e ) of the immunohistochemical staining for the P2rx1 protein (original magnification ×200, scale bar = 200 μm). All quantification results are presented as mean ± SEM. *p

    Journal: Scientific Reports

    Article Title: Induction of detrusor underactivity by extensive vascular endothelial damages of iliac arteries in a rat model and its pathophysiology in the genetic levels

    doi: 10.1038/s41598-019-52811-4

    Figure Lengend Snippet: Repression of P2rx1 by progressive vascular endothelial damage. ( a – c ) RQ-PCR ( a ) and western blot ( b , c ) analysis of the expressions of the cholinergic receptor muscarinic 2 ( Chrm2 ), cholinergic receptor muscarinic 3 ( Chrm3 ), and purinergic receptor P2X 1 ( P2rx1 ) genes in the bladders of rats in the indicated groups. Expression levels of the indicated transcripts are presented as % Gapdh . For western blot analysis, β-actin was used as a loading control, and the expression levels of the indicated proteins were normalized to the β-actin value. ( d , e ) Representative images ( d ) and quantification analysis ( e ) of the immunohistochemical staining for the P2rx1 protein (original magnification ×200, scale bar = 200 μm). All quantification results are presented as mean ± SEM. *p

    Article Snippet: Protein levels were assessed from 50 μg of extracts separated using 10% SDS-PAGE gels by probing with antibodies specific to P2rx1 (APR-001, Alomone Labs), Chrm2 (AMR-002, Alomone Labs,), Chrm3 (AMR-006, Alomone Labs, Israel), α-SMA (ab5694, Rabbit, Abcam), vimentin (sc-6260, Santa Cruz Biotechnology Inc., CA), and β-actin (Sigma-Aldrich).

    Techniques: Polymerase Chain Reaction, Western Blot, Expressing, Immunohistochemistry, Staining