An equal amount of lysate (20 µg) was loaded and the blots were probed with a peptide antibody directed to the C-terminus of HsPCFT. The eastern blot, or eastern blotting, is a biochemical technique used to analyze protein post-translational modifications (PTM) including the addition of lipids, phosphates, and glycoconjugates. Keywords: Signal transduction, Glycosylation, O-GlcNAc, Method(s), Detection, Analysis, Galactosyltransferase, O-linked, Glycosylation INTRODUCTION The modification of Ser and Thr residues by monosaccharides of O-linked β-N-acetylglucosamine (O-GlcNAc) is a common, dynamic, and essential modification of nuclear and cytoplasmic proteins. They have done it in rat brain homogenate.
The protein can then be analyzed by SDS-PAGE and protein staining or Western blotting to detect the protein of interest. Flagella are required for biofilm formation, as well as adhesion to and invasion of epithelial cells. In general, a sample is either left untreated or treated with the enzyme/chemical. (Predicted to have 67 O-linked glycosylation sites + 8 N-linked glycosylation sites). It is glycosylated at 4 amino acids. Recognition of flagellin via the Toll-like receptor 5 (TLR5) contributes to exacerbate B. cenocepacia … Lower panel; Western blot analysis of wild-type and glycosylation mutant HsPCFT membrane proteins. N-linked glycosylation sites are estimated to be 2.5 kDa extra per site to the total weight = 20 kDa more. Western blot analysis using the PD-L1 extracellluar domain (ECD)- or intracellular domain-specific antibodies indicated that PD-L1 glycosylation occurred on … The Western blot assay provides valuable information about a protein including abundance, the apparent molecular mass, post-translational modifications and splice variants. Burkholderia cenocepacia is an opportunistic pathogen threatening patients with cystic fibrosis. I am doing western blotting for a protein(mol weight 77kD). Deglycosylating enzymes and chemicals can be used in initial studies to study glycosylation of a protein. Approximately half of all proteins typically expressed in a cell undergo this modification, which entails the covalent addition of sugar moieties to specific amino acids. Company spec sheet shows a band at 100kD. However, analysis of the protein can be difficult if multiple bands appear on the blot. Data are the mean ± SEM from three independent experiments. Glycosylation is a critical function of the biosynthetic-secretory pathway in the endoplasmic reticulum (ER) and Golgi apparatus.