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    Alomone Labs anti machr m2
    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

    Expression of key components in the mAChR signaling pathway. The expression levels of (A) mAChR M2, and (B) mAChR M3 and their respective downstream proteins were determined by (a-c) western blot and (d) immunofluorescence assays. Band intensities were evaluated relative to the intensity of the actin bands. Data are presented as mean ± standard deviation of three replicates (n=5-6 rats per treatment group). * P

    Journal: International Journal of Molecular Medicine

    Article Title: Spicatoside A in red Liriope platyphylla displays a laxative effect in a constipation rat model via regulating mAChRs and ER stress signaling

    doi: 10.3892/ijmm.2018.3960

    Figure Lengend Snippet: Expression of key components in the mAChR signaling pathway. The expression levels of (A) mAChR M2, and (B) mAChR M3 and their respective downstream proteins were determined by (a-c) western blot and (d) immunofluorescence assays. Band intensities were evaluated relative to the intensity of the actin bands. Data are presented as mean ± standard deviation of three replicates (n=5-6 rats per treatment group). * P

    Article Snippet: Following the separation of total proteins (30 μ g) by 4-20% SDS-PAGE, these proteins were then transferred to nitrocellulose membranes for 2-3 h at 40 V. Subsequently, the membranes were incubated with primary antibodies targeting mAChR M2 (cat. no. AMR-002; 1:1,000; Alomone Labs, Jerusalem, Israel), mAChR M3 (1:1,000, cat. no. AMR-006; Alomone Labs), phosphoinositide 3-kinase (PI3K, 1:1,000, cat. no. 4292S; Cell Signaling Technology, Inc., Danvers, MA, USA), phosphorylated (p-) PI3K (1:1,000, cat. no. 4228S; Cell Signaling Technology, Inc.), protein kinase C (PKC, 1:1,000, cat. no. 2058S; Cell Signaling Technology, Inc.), p-PKC (1:1,000, cat. no. 9376S; Cell Signaling Technology, Inc.), inositol-requiring enzyme (IRE) 1α (1:1,000, cat. no. ab37073; Abcam, Cambridge, UK), IRE1β (1:1,000, cat. no. SC-10511; Santa Cruz Biotechnology, Inc., Dallas, TX, USA), p-IRE1 (1:1,000, cat. no. 48187; Abcam), c-Jun N-terminal kinase (JNK, 1:1,000, cat. no. 9252; Cell Signaling Technology, Inc.), p-JNK (1:1,000, cat. no. 9251; Cell Signaling Technology, Inc.), guanine nucleotide binding protein (G) α (1:1,000, cat. no. ab128900; Abcam), eukaryotic initiation factor (eIF) 2α (1:1,000, cat. no. 9722; Cell Signaling Technology, Inc.), p-eIF2α (1:1,000, cat. no. 9721; Cell Signaling Technology, Inc.) or β-actin (1:3,000, cat. no. A5316; Sigma-Aldrich Co.; Merck KGaA, Darmstadt, Germany) overnight at 4°C.

    Techniques: Expressing, Western Blot, Immunofluorescence, Standard Deviation