The secondary antibody would the overlap the primary antibody by binding to it. The secondary antibody (anti-rabbit HRP labelled) used is conjugated with an enzyme Horse radish peroxidase which then binds to its substrate tetramethyl-benzidine (TMB) to produce a blue color indicating the presence of lysozyme. SDS-PAGE gel electrophoresis is a process which separates proteins based on molecular weight. The purpose of this method is to separate out the lysozyme by its weight. The weight is known to be 14.6Da.
The presence of the black precipitate indicates that sulfide and not sulfate, which gives off a brown color, was present in the reaction. Precipitation reactions. Precipitation reactions involve the denaturation of proteins. This is so because the most common observation with protein denaturation is the coagulation or precipitation of the product. Denaturation is the disruption and/or the possible destruction of the secondary and tertiary structures in the protein.
Each solution was then plated on an LB/kanamycin/IPTG plate. Additionally transformation #2 was plated on a LB/kanamycin plate, and transformation #4 was plated on an LB plate. The purpose of including kanamycin on the plates was to selectively screen for plasmid containing E. Coli cells. The IPTG was to selectively screen for E. Coli cells with the plasmid containing the EGFP insert. The plates were then incubated over night at 37° C. DNA Purification The objective of this experiment was to isolate plasmid DNA.
To calculate RMSd we used PTRAJ from the AMBER package and then we compared the conformation of the enzyme after the simulations at the 0.1 ps interval.  2.8.2. The Hydrogen Bond Analysis Hydrogen bonds formed between residues of the protease and between residues and water molecules, were analyzed using PTRAJ. Only bonds with a distance less than 3.5 Å with the angle of interaction greater than 140 ° were considered. The output file gave us the results which hydrogen bonds were formed, with which occupancy, distance and angle.
Western blotting is a procedure used to detect specific proteins in a given sample. Gel electrophoresis is used to separate the proteins which can be observed as thick and thin bands on the electrophoresis gel. In this experiment we use SDS-free polyacrylamide gel. Sample proteins used in this case are bovine serum, human serum, goat serum, chicken serum and horse serum. Since the SDS is negatively charged, sample proteins move to the positively charged anode through the gel.
ABSTRACT NRC-04, a novel antimicrobial peptide derived from skin mucous secretions of flat fish winter flounder, shows a broad spectrum of antimicrobial activity. In order to understand the conformational change of NRC-04 in different types of membrane, our team did experiments on NRC-04 with negatively charged bacterial surface membrane mimetic micelles sodium dodecyl sulphate(SDS), zwitterionic eukaryotic middle membrane mimetic micelles dodecylphosphocholine(DPC), gram-negative bacteria outer membrane mimetic micelles Lipopolysaccharide(LPS) and bacterial inner membrane mimetic micelles 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol(POPG). Fluorescence test shows that the C-terminus tryptophan residue of NRC-04 interacts with the hydrophobic
Fig 1 represents the vein diagram or the differentially expressing proteins from MCF-7 using Venny (http://bioinfogp.cnb.csic.es/tools/venny/index html).1628 proteins were identified from our proteomics data among which 403 proteins were differentially expressed in both samples. Subsequent filtering by applying statistical criteria of a 95% up- or down-regulation likelihood ( [p > 1] 1] > 0.95) and a fold-change higher than 30% (ratio of either 1.3) as significantly altered levels of expression. 212 proteins were identified as up-regulated and 191 proteins as down-regulated in progesterone treated samples compared to control. The complete list of proteins and their relative quantities are given in the supplementary file No.1. Among the total protein samples, some were unique to only control while few others unique to different time points of treatment, and many others overlapped.
Cycloheximide applies its impact by interfering with the translocation steps in protein synthesize (development of two tRNA atoms and mRNA in connection to the ribosome), hence blocking translational prolongation. Cycloheximide is generally utilized as a part of biomedical research to repress protein synthesize in eukaryotic cells except for S.aureus and E.coli contemplated in vitro (i.e. outside of microorganism). It is cheap and works quickly. Actually after the interaction of 72 hours, both growth of E.coli and S.aureus will be inhibited by Cycloheximide antibiotic.
OBJECTIVE The objective of this experiment was to analyze and determine the unknown concentration of a protein solution by utilizing two different colorimetric techniques; Biuret and Lowry. The Biuret method was used with unknown #2 and the Lowery method was used for unknown #1. After the concentration of each unknown was analyzed (by Biuret or Lowry method), the alternate objective was to compare the results achieved by each method and to determine if the results from the approaches were consistent in contrast to each other. THEORY Biuret and Lowry methods use colorimetry as a tool to analyze protein concentration quantitatively. While both methods are respected and widely used, the Lowry method has a sensitivity level that is approximately 100 times greater than the Biuret method.
CHAPTER I INTRODUCTION 1.1 RESEARCH BACKGROUND This research is about the extraction of keratin protein from chicken feathers using strong fluid extraction process. This extraction procedure utilized a dissolvable as lessening specialists to separate salt linkages, disulphide bond and hydrogen bond of the keratin fiber to break down into protein. Thus, it will likewise diminish the steadiness of keratin strands in the strong structure found in feathers. Proteins are polymer formed by one or more long chains of amino acid residues that are capable of promoting intra- and inter-molecular bonds so that it allows the resultant materials to have a large variation in their functional properties. Amino acids linked together by peptide bonds, form a polypeptide chain.