Human/Mouse/Rat MuRF1/TRIM63 Antibody

Detection of Human/Mouse/Rat MuRF1 by Western Blot. Western blot shows lysates of human heart, mouse heart, and rat muscle tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse/Rat MuRF1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5366) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for MuRF1 at approximately 41-44 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

Detection of Human MuRF1/TRIM63 by Simple Western^TM. Simple Western lane view shows lysates of human heart tissue, loaded at 0.2 mg/mL. A specific band was detected for MuRF1/TRIM63 at approximately 46 kDa (as indicated) using 10 µg/mL of Goat Anti-Human/Mouse/Rat MuRF1/TRIM63 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5366) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of Zebrafish MuRF1/TRIM63 by Western Blot Blocking MuRF1-mediated proteasome degradation preserves myofibril integrity in tre/ncx1 deficient hearts.(A) Z-lines were visualized by alpha -actinin staining. By 72 hpf, sarcomeres are disassembled in hearts of uninjected (tre) and control morpholino-injected (tre +ctlMO) tremblor embryos. Murf1a/murf1 b knockdown does not affect sarcomere integrity in wild type embryos (WT +MO), but prevents sarcomere degradation in tre (tre +MO). Similarly, treatment with a proteasome inhibitor, MG132, preserves myofibril integrity in tre cardiomyocytes (tre +MG132). Scale bar, 10 μm. (B) Graph shows % of embryos with periodic alpha -actinin staining at 72 hpf. (C) Western blot detecting MuRF1 and beta -actin proteins in uninjected control (WT control) and murf1a/murf1 b knockdown (MuRF1 MO) embryos. Chi-squared test, *p<0.05; **p<0.01; ***p<0.001. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28826496), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse MuRF1/TRIM63 by Western Blot The qPCR (quantitative real-time PCR) (A) and Western blot analysis (B,C) showed the mRNA (A) and protein (B,C) expressions of TRAF6, MAFBx, and MuRF1 in the TA muscle of mice injected with vehicle (saline, control) or Dex (dexamethasone sodium phosphate in saline, 10 mg/kg) respectively. Data are presented as mean ± SD, n = 9 per animal group, * p < 0.05 versus control. Also shown (B) is a representative Western blot image. GAPDH and tubulin were used as a loading control in qPCR and Western blot analysis. Image collected and cropped by CiteAb from the following publication (https://www.mdpi.com/1422-0067/15/6/11126), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse MuRF1/TRIM63 by Western Blot (A) Micrograph showed the morphology of C2C12 myotubes cultured in vehicle (0.1% ethanol-containing plain medium, control) or stimulated by Dex (dexamethasone in vehicle) respectively. Scale bar = 100 μm; (B) Bar graph compared the diameter of C2C12 myotubes cultured in vehicle (control) or stimulated by Dex respectively; and (C,D) Representative Western blot image and Bar graphs displayed the protein expression of TRAF6, MAFBx, MuRF1, and desmin in C2C12 myotubes cultured in vehicle (control) or stimulated by Dex respectively. Tubulin were used as a loading control in Western blot analysis. All data in bar graphs are presented as mean ± SEM (standard error of the mean) from three independent experiments, * p < 0.05 versus control. Image collected and cropped by CiteAb from the following publication (https://www.mdpi.com/1422-0067/15/6/11126), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse MuRF1/TRIM63 by Western Blot The qPCR (A) and Western blot analysis (B,C) showed that C2C12 myotubes were transfected with TRAF6-siRNA or control-siRNA; Micrographs (D) showed the morphology of Dex- or vehicle-treated C2C12 myotubes after transfection with TRAF6-siRNA and control-siRNA respectively. Scale bar = 100 μm; Bar graph (E) showed the diameter of Dex- or vehicle-treated C2C12 myotubes after transfection with TRAF6-siRNA and control-siRNA respectively; The qPCR and Western blot analysis showed the mRNA (F) and protein (G,H) expressions of TRAF6, MAFBx, and MuRF1, as well as the protein expression of pFOXO-1 in Dex- or vehicle-treated C2C12 myotubes after transfection with TRAF6-siRNA and control-siRNA respectively. GAPDH and tubulin were used as a loading control in qPCR and Western blot analysis. All data in bar graphs are presented as mean ± SEM from three independent experiments, * p < 0.05. Image collected and cropped by CiteAb from the following publication (https://www.mdpi.com/1422-0067/15/6/11126), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse MuRF1/TRIM63 by Western Blot (A) Masson trichrome staining image of the TA muscle from mice co-injected with both Dex and control-siRNA, with both Dex and TRAF6-siRNA, with both vehicle and control-siRNA, or with both vehicle and TRAF6-siRNA, respectively, for 14 days; (B,C) Bar graphs showed the weight (B) and cross sectional area (CSA, C) of the TA muscle from mice co-injected with each of the above four combinations respectively. Scale bar =20 μm; (D–F) The qPCR and Western blot analysis showed the comparison in the mRNA (D) and protein; and (E,F) expression of TRAF6, MAFBx, and MuRF1, as well as the protein expression of pFOXO-1 in the TA muscle of mice co-injected with each of the above four combinations respectively. GAPDH and tubulin were used as a loading control in qPCR and Western blot analysis, respectively. All data in bar graphs are presented as mean ± SD, n = 9 per animal group,* p < 0.05. Image collected and cropped by CiteAb from the following publication (https://www.mdpi.com/1422-0067/15/6/11126), licensed under a CC-BY license. Not internally tested by R&D Systems.