Coomassie

SDS page Name: Shelby Clark Lab Partners: Kaliah Goodman, Howsikan Kugathasan, and Suntesia Bowen Date of Laboratory: September 21, 2016 Goal of Laboratory: The goal of this laboratory was to successfully make a gel and to run an SDS-PAGE and determine molecular weight (MW) of an unknown protein sample by comparing it to Log10 of the migration of molecular weight of the standards. Background: The purpose of SDS-PAGE is to separate proteins according to their size, and no other physical feature

Bradford Assay on Unknown Concentrations of Proteins Taylor Coleman September 27, 2016 Lab Group 3 BIOL 1111: General Biology Lab Fall 2015 Section 107 Chad Perry Abstract Colorimetric assay is a process of determining a concentration of a solution based on absorbance of light. The purpose of this lab is to determine if the Bradford assay is an accurate way to determine an unknown concentration of two samples of protein. The Bradford assay is done by measuring wavelength of light passing

change in the solution depending on the amount of peptide or amino acid reactions with the reagents. This color change can then be measured using a device called a spectrophotometer. The Bradford assay is a one- step process, which involves the Coomassie dye reacting with the amino acids arginine and lysine. It under goes changings in acidic conditions

performed in order to confirm the proteinuria and to estimate the total amount of protein (4). Another method available for detecting proteinuria is called the Bradford assay. This quantitative method is based on the binding of proteins with the dye Coomassie Blue which increases the absorption of the dye at 595nm. The absorbance measured by spectrophotometry is related to the amount of proteins present in the

Title - Gel Filtration and SDS Polyacrylamide Gel Electrophoresis (P.A.G.E) Gel Filtration, SDS-PAGE Objective – The purpose of this lab was to separate a mixture of Hemoglobin, BSA, Blue Dextran, and yellow food coloring by size into fractions using gel filtration. Also simple non-quantitative assay was performed to determine the fractions that contained proteins. Another objective was to analyze the protein content of being BSA or Hemoglobin and estimate the molecular weight of the proteins collected

For this experiment, whole bovine blood was used. The first process was to separate the blood into cellular and plasma fractions. 100 µL of whole bovine blood was transferred into a yellow microcentrifuge tube that was labeled WB using a P-200. 50 µL of whole blood was added to a blue microcentrifuge tube labeled WB. Both tubes were capped and placed in ice. 2 mL of the remaining blood was transferred into a Clear 2 mL tube using a P-2000 and centrifuged for 5 minutes at 8000 RPM. Afterward, 800

MATERIALS AND METHODS The BIORAD dye was firstly prepared by dissolving 0.225g of Coomassie Brilliant Blue G-250 (Merck, South Africa), in 67.5ml of Methanol (Riedel-de-Haёn, Germany) and 291ml of 85% (w/v) Phosphoric Acid, diluted to 450ml with dH2O. Six test tubes were labelled SC1-6 and the 0.1mg/ml stock of Bovine Serum Albumin (Roche

The incorporation a plasmid vector, pGLO, into Escherichia coli via transformation is described. This recombinant plasmid that has been altered contains the ability to fluoresce green under ultraviolet light because of the Green Fluorescent Protein (GFP). pGLO also encompasses an antibiotic resistance gene, bla, that allows E. coli to grow in media that contains penicillin derivative antibiotics such as ampicillin. We conclude that the protein, GFP, is indeed responsible for the green fluorescence

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.

ion-exchange chromatography method via a cation (CM) or anion (DEAE) exchanger. This was followed by a SDS-PAGE gel electrophoresis technique to determine the original mixture’s proteins and concentration with the help of a Bradford assay, which utilizes a Coomassie dye to bind to proteins. After all techniques were performed, Protein 1 was found to have an absorbance value of 0.451 and a concentration of 0.019 µg/µL. Protein 2 was found to have an absorbance of 0.373 and a concentration of 0.016 µg/µL. Ultimately