Mouse Legumain/Asparaginyl Endopeptidase Antibody

Detection of Mouse Legumain/Asparaginyl Endopeptidase by Western Blot.

Western blot shows lysates of J774A.1 mouse reticulum cell sarcoma macrophage cell line and Neuro-2A mouse neuroblastoma cell line. PVDF membrane was probed with 0.2 µg/mL of Sheep Anti-Mouse Legumain/Asparaginyl Endopeptidase Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2058) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). Specific bands were detected for Legumain/Asparaginyl Endopeptidase at approximately 37 and 56 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Legumain/Asparaginyl Endopeptidase by Western Blot

The presence of VDAC1-DC in ccRCC cells decreases or abolishes ciliation. A, Triple immunofluorescence labeling and merged images with acetylated alpha-tubulin (Acet. alpha-tubulin in red), Arl13b (in green) and DAPI (in blue). B, Electron microscopy of RCC4+pVHL cells. C, Quantitative analysis of the ciliation percentage was assessed by confocal fluorescence microscopy (n=100-300 cells). D, Both cell lines were seeded at the same density and incubated in Nx for 48h with or without serum. Percentage of ciliated cells, proliferation and FACS analysis were measured. The mean ± SEM is representative of three independent experiments carried out in duplicate. E, F and G, RCC4 cells were transfected with control siRNA (siCtl), (E) siHIF-1 alpha, siHIF-2 alpha and siHIF-1/2 alpha, (F) siVDAC1 and (G) siLGMN. Cell lysates were analyzed by immunoblotting for HIF-1 alpha, HIF-2 alpha, VDAC1, LGMN and beta- tubulin/Actin or HSP90 were used as a loading control. Quantitative analysis of the ciliation percentage was assessed by confocal fluorescence microscopy (n=100-300 cells). A * p<0.05 shows significant differences. Quantification of VDAC1 and VDAC1-delta C protein levels (E). Experiments have been proceeded without serum. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/32194829), licensed under a CC-BY license. Not internally tested by R&D Systems.