The Use Of Sodium Dodecyl Sulphate ( Sds ) Polyacrylamide Gel Electrophoresis Essay

This experiment will be done as the first step in determining an unknown protein. The second part of the protein determination will be done using the Western blot technique. We will use the Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) method to separate proteins by their molecular weight. This method is often used to determine the purity and weight of a protein. Electrophoresis is a method of separating proteins based on their chemical and physical properties. Gel electrophoresis has a gel support mediums that includes polyacrylamide, starch and agarose. The gel is chemically inert, it is easy to handle, and can me made to fit a desirable porosity. This makes it good to use for molecules with high molecular weights. After separating the gel, it will be stained with Coomassie Blue to visualize the protein.

The IgG antibody was separated by using AKTA FPLC by setting the method as:- Column- HiTrap protein G HP 1 mL; Column pressure limit- 0.300 MPa; Flow rate- 1 mL/minute; Sample injection- 0.200 mL; Loading buffer, A- 0.05 M sodium phosphate, 0.15M NaCl, pH 7.0; Elution buffer, B- 0.1 M glycine-HCl, pH 2.7; Neutralization buffer, C- 1 M Tris-HCl, pH 9.0. Different column volume for individual step was then set as- 1 CV 0% B wash and equilibration step; 1 CV 0% B loading; 4 CV 0% B washing; 10 CV 100% B eluting; 5 CV 0% B equilibration. After setting the method, the system was washed with buffer B at the flow rate of

ELISA works on the principle of an antigen binding to specific antibody (lock and key), which can be used as a way to identify quantities of proteins in a small sample of fluid. The specific proteins used in an ELISA are estimated quantitatively. The ELISA test is carried out by incubating the serum that contains the antigen of interest with antibody’s within a well, in order for the antibody’s to bind with the specific antigens. The plate is then washed with a mild detergent in order to remove any proteins that have not been bound. The washing of the plates is carried out between every step in order

The testing of various proteins was performed by comparing the molecular weight of proteins using SDS PAGE. The molecular focus in the lab was the testing of proteins, which are macromolecules consisting of amino acid monomers linked through chemical bonds. These proteins have a hierarchy of structure that consists of folding that determines the direct function of each protein.. The molecular weight of these proteins were measured using SDS PAGE. SDS PAGE stands for sodium dodecylsulfate polyacrylamide gel electrophoresis. SDS is an anion detergent that binds with the protein structures and causes them to separate due to the change in bonding charge. SDS and heat are how the proteins are denatured. The process of denaturing a protein is breaking

SDS-PAGE separates proteins according to their size. Sodium dodecyl sulfate (SDS) dissolves hydrophobic molecules and carries a negative charge. A cell incubated with SDS would have its membranes dissolve and all the proteins becoming soluble and covered with negative charges. As a result, all the proteins only have their primary structures and a large negative charge allowing them to migrate towards the positive pole of an electric field. Polyacylamide gel is used to separate proteins according to their sizes. The gel has pores of different sizes that act as obstacles for the proteins to pass through. Switch off the current, stain the proteins, and the end result would be bands of protein separated according to their molecular weight. One

In this experiment, the erythrocyte membranes were dissolved in sodium dodecyl sulfate (SDS), which subsequently led the proteins to resultantly denature and separate from the lipids. The samples are then placed in the apparatus which carries out electrophoresis; the mixtures are subject to an electric field (the SDS places a greatly negative charge on the polypeptides, making their intrinsic charge irrelevant) and the lighter ones migrate faster than heavier ones. Subsequently, Coomassie blue is added so that the polypeptides can be observed on the polyacrylamide gel (blue lines will indicate the polypeptides). However, some components (the heavily glycosylated glycoproteins) of the experiment could not be stained through Coomassie blue or through other normally used protein staining techniques; luckily, they can still be made visible by the periodic acid-Schiff (PAS) staining

Gel Electrophoresis is a technique used to separate macromolecules fragments like DNA and protein by their size. Also, the proteins can be separated according to their size and their charge of different proteins with the different charges. Therefore, the material being separated and is placed into a gel-like substance called agarose gel. Because each DNA molecule is negatively charged, it can be pulled through the gel by an electric field. Small DNA molecules move more quickly through the gel than larger DNA molecules. (Biotechnology) For example, the DNA are separated to get a DNA fingerprint for forensic purposes, to get a DNA fingerprint for paternity testing, to check a PCR reaction, and to get a DNA fingerprint so that you can look for

Gel Electrophoresis is used to separate macromolecules like DNA and RNA by their size, or proteins by their charge and size. In this experiment, I used the Gel electrophoresis to determine the presence of all dyes in a specific dyes mixture.

Report 3 could have been improved by talking about what sodium carbonate and sodium bicarbonate is in the introduction as well as removing the lists under Materials and Methods. The Materials and Methods should always be in sentence form in the past tense. Also more than two trials should have been done because replication is very important to scientific experiments. This ensures that there are no outliers in the results. Also, the author should have avoided personal pronouns like “I” in the conclusion. A sentence that could replace the last would be something like “Because the actual percent composition of the mixture is unknown, the accuracy of the current

DNA is commonly used in the modern legal system to release the innocent or imprison the guilty. There are two different ways to create DNA evidence through science technology, PCR and gel electrophoresis. Polymerase Chain Reaction, PCR, is used to create thousands to millions of copies of DNA, out of a small piece of DNA, this piece is used to make a DNA profile. Doing this makes it easier to test the DNA. PCR is used to find if a gene has pathogens. They are an infectious agent that creates illness or disease and it’s commonly a virus, parasite or bacteria. (1) Gel electrophoresis is the technique used to separate molecules from DNA, RNA or proteins, based on the size or electric charge. It can be used in DNA fingerprinting, the findings and purification of nucleic acids and proteins. Gel electrophoresis can be used to find pathogens that can be found in blood, tissues or sources like food. (2) These are some of the methods that are used in court and police cases to discover DNA.

Gel electrophoresis has had a very big impact on the world of science today. In this present day, scientists now have a simple and definite way to test and contrast a wide variety of different DNA samples, leading to more complex and particular DNA testing (LinkedIn Corporation, 2017). Gel electrophoresis is now a scientific method in which is an absolute necessity in science, particularly in the forensics field. This process of gel electrophoresis gives forensic scientists the ability to retrieve an indisputable link to a person of suspect or anybody for that matter.

Thank you for the opportunity for my first journal peer review. After I had submitted, I saw that I had some grammatical mistakes myself. However, please feel free to pass on any feedback on my review so that I can better serve the journal in the future, if my services are needed again.

The peer review is very helpful. Alina suggested I provide more details and explanations to support my argument. I agree

From peer reviewer #1’s comments, I accept that I should have written out the full name of S. marcescens the first time I mentioned it. I also accept their comments that I should add a title and key words to my paper, as well as switch the order of the references and appendix. Also, as peer reviewer #1 suggested, I added all of the required elements to my appendix, including the null and alternate hypothesis as well as the interpretation based on the p-value. I also changed my in-text citations so that they are in accordance with the CSE format. I reject the reviewer’s comments that my method’s section was too detailed because in some other research papers that I have read, the methods section was equally verbose. I also reject the reviewer’s comment that I would remove the

There are several potential explanations for why our experiment did not yield significant results in support of our