Lipoprotein isolation and characterization: Leghon red hens commercial eggs ( > 73g.) were used for this study. The shell was broken, the white of an egg was eliminated and the yolk was put on a filter paper to roll in it to eliminate completely the remainders of the white adhered. The yolk membrane was punctured and the content of the yolk (approximately 15 ml) was deposited in a test tube. An equivalent volume of saline solution was added (NaCl 0.16 M; pH = 7) and it was mixed completely. The resultant solution was centrifugated for 30 minutes at 9000 rpm at 4ºC in a rotor Beckman JA-20, eliminating the pellet. The supernatant was collected to obtain the lipoproteins by gradient density ultracentrifugation: 10ml of the yolk solution were taken and then 0.9 g of potassium bromide (KBr) was added. The bromide was dissolved with a smooth agitation, with care not to denature the proteins. Straight afterwards, saline solution was added (NaCl 0.16 M, pH=7) to the 10ml of the yolk solution with a Pasteur pipette, avoiding the sample diffusion, forming two phases and filling the tube completely. The ultracentrifugation was carried out during 19.5 h at 4ºC and 45000 rpm. (244.500 x g) in a Kontron Centrikon T-2190 ultracentrifuge in a TFT 50.38 rotor, …show more content…
Their visualisation was performed using Coomassie brilliant blue (G-250). GS-800TM Calibrated Densitometer (BioRad, California, USA) was used to scan the gels and the resulting image file was assayed using the Quantity One analysing software (BioRad, California, USA). Molecular masses were estimated according to Precision Plus Protein unstained standards (BioRad, California, USA) which contains ten highly purified recombinant proteins with molecular masses from 10 to 250
Figure 20 and 21 casting tray Figure 22 gel box After 1 hr., take out the gel from gel box carefully, place it into the machine, so that the DNA gel electrophoresis can be visualize under UV light. Figure 23 gel documentation system, used to visualize gel electrophoresis with UV light NMR spectroscopy First of all, 3 samples were prepared, peptide in SDS, DPC, and buffer. The sample temperature was maintained at 298 K, prepared by supervisor and H(hydrogen) in SDS and DPC micelle was replaced with D(deuterium) , so that in proton NMR, peptide won’t be interfered by H in micelle. amount of peptide in sds and dpc
To limit the amount of errors or contamination in any procedure lab safety rules, gloves, and the aseptic technique was strictly enforced throughout the experiment. The first step to identifying the unknown bacterium was the Streak Plate Method. This method is used to isolate a pure culture from a mixed culture. Also, this method included streaking a tryptic soy agar (TSA) plate into four quadrants, and later incubating the plate for 24 hours.
During this experiment we are going to test this hypothesis by putting the egg into vinegar so the shell disintegrates and we get a permeable plasma membrane. After we will place it in two different substances one being corn syrup and the other is water with food coloring. We will observe how osmosis moves water across the plasma membrane during this whole experiment and how passive transport works. We will also observe Hypertonic and Hypotonic environments and how they move water across the plasma membrane going from high to low
In the last 24 hours, I had an egg sandwich. The brand of eggs that I consumed was Egg Land’s Best. The competitive strategy that Egg-Lands Best utilizes is Broad Differentiation strategy. They command a premium price for their product. For example, the cost of Egg-Lands best eggs can range from $2.67 and up compared to competitor’s prices which can be as low as $1.00.
The egg-speriment was one of the ways to observe how the cell membrane works with different liquids entering and leaving. The egg is a model of the cell membrane that is soaked in vinegar for Days 1 and 2, water for Days 3 and 4, food coloring with water for Day 5, salt water for Day 6, and finally Arizona Tea for Day 7. My hypothesis was when the egg is soaked in vinegar, it will make the shell rougher, water will make it soft, food coloring will make it colorful, and salt water will make it a rocky shell. In this egg-speriment, the manipulated variables is the liquids, the responding variable is circumference, and the controlled variable is the egg.
Egg Drop Activity was one of my favorite experience that I was excited to explore in this class. while working on this activity, I had a fun time to cooperate with my classmate, Briana to try different ways to protect the egg. However, I was struggling to layering materials provided to protect the egg because I needed to hold the egg carefully. While exploring with this activity, I learned that from this experience that everybody had different techniques, and they also used diverse materials to protect the egg. Personally, I took a long time to think critically and creatively to make the protection for the egg.
Abstract The purpose of this experiment is to test for mitochondrial activity by isolating different organelles using the differential centrifugation process. Studying mitochondria is extremely important because they control the death and life of the cell by regulating the apoptotic signals (Frezza et al 2007). Also they are responsible for the metabolic reactions (aerobic respiration) and the production of ATP (Frezza et al 2007). Three hypotheses were formed based on my knowledge.
Genius When using gas in the egg drop challenge, it spreads the amount of energy transferred from the ground to the egg. Making the energy from the ground to the egg less, because if you look closely at a balloon filled with air. When it hits the ground, the gas in the balloon spreads out inside the balloon, and then contracts back. The solid object (the egg) on top of the balloon will sink into the balloon, because it’s a solid and heavier than the gas in the balloon. Let’s say you have an egg that weighs about one pound, you have to build a sculpture that slows down something moving at 9.8 meters per second.
Starch solution is then placed into the test tube at a quantity of 5 mL. 5 drops of Lugol’s Iodine solution is added to the test tube. If the color changes, then it is known that starches are present in the solution. Proteins are next tested. In order to do this, 5 mL of gelatin solution is added to the test tube. 10 drops of Biuret’s reagent are added to test for protein.
The investigation was carried out to identify the presence or absence of biological molecules in serum 2216. If the concentration in each test tube of the dilutions carried out will be more concentrated then the concentration of the test tube before it, then the color will be at an equal concentration with the other dilutions performed. The hypothesis was wrong because of the difference in concentrations due to the different measurements within the dilutions done. The test for starch was to add a drop of iodine solution to the pipette in the spotting tile. A reducing sugar solutions is add inside a test tube with 3 drops to then add 3 drops of benedicts and plane in a water bath.
The eggs were placed in the solutions, charting the change in weight after 15, 30 and 45 minutes; handling the eggs carefully each time and wiping off any excess water to avoid a misread by the scale. It is necessary to remove the egg and weigh individually each time to get the most accurate results. After the experiment was done, the eggs were placed back in their containers and the solution discarded of. It is imperative not to put the solution back in the containers.
1.1 Abstract The purpose of quantitative analysis of protein using a spectrophotometer is to measure the concentration of proteins in a given sample. The experiment is conducted by laboratory method (Biuret Test) and using spectrophotometer to analyze the absorbance of reactants at 540 nm, hence determining the concentration of the proteins in a given sample. The purpose of stopped enzyme assay to study B-galactosidase is to determine the effect of temperature and concentrations of substrate on enzyme activity.
The preset study mainly focuses on biomarker Lipocalin-2 and
Finally, the amplified DNA regions are compare using a gel. DNA Profiling
The molecular weight of this glycoprotein is about 65,000. This protein is similar to the conglycinin from soy proteins with the major IgE epitopes within this extension region. Ara h 1 is a protein with high thermal stability but showed minor structural changes in 5M urea. It has also been observed that few of the IgE binding epitopes of Ara h1 are resistant to pepsin degradation [22].