Biochemistry 101

"There are several ways of estimating the protein concentration such as amino acid analysis following acid hydrolysis of the protein; analyzing the changes in the spectral properties of certain dyes in the presence of proteins; and spectrophotometric estimation of the proteins in near or far UV region. Although dye-binding assays and amino acid analysis following acid hydrolysis of the protein can be used for estimating the protein concentration for both pure as well as an unknown mixture of proteins; UV spectroscopic quantitation holds good for the pure proteins. If a protein is pure, UV spectroscopic quantitation is the method of choice because it is easy and less time consuming to perform; furthermore, the protein sample can be recovered back". One of the biggest obstacles as an overall whole, was the calculation part of the experiment, I have attached a video that helps better explains the process of finding the standard curve.  REFERENCES [1] Scopes, R. K. (1974) Measure...

In this specific lab, we used a purification method,  Gel filtration chromatography, to separate a mixture of two molecules known as Blue Dextran and Fluorescein based on the size of the molecule. A column was prepared using a gel filtration medium called Sephadex G-50. Prior to class Dr.Regmi made an elution buffer of Na2HPO4/ NaH2PO4  pH 7.0 for size-exclusion chromatography. Through the duration of this lab, I  learned that the atoms that are bigger in size normally elute the section first before more modest particles can, due to their exchanges with the fixed stage, the beads. Below is a list of a variety diameters of organelles; this determines the RCF required Below is a video of the Gel-filtration Sephadex G-50 Procedure

In the third lab experiment I was able to isolate the Nuclei and Mitochondria from chicken liver using differential centrifugation. Though this was my first time ever using a centrifuge, I was able to learn and understand the theory behind the process. This technique is very common and was used in the experiment to separate to major organelles, the nucleus and mitochondria,  through a stepwise increase in centrifugation speed. Chicken liver and the cold fractionation buffer was homogenized and filtrated in centrifuge tubes. We used two different settings of RPM. 2500 RPM for the nuclei and 5600 for the mitochondria due to the size of the diameter, the smaller the particle the greater the RCF.  Below is a video of the process of Cell Fractionation. . .

I n 1908, Lawrence Joseph Henderson formulated an equation which explained carbonic acid as a natural buffer. Another gentlemen , by the name of Karl Albert Hasselbalch later used Henderson's formula to work a logarithmically relation, that measures pH derivation. A buffer is simply a solution that resists changes in pH due to amounts of a base and acid as well as dilution .  Buffers are essential to everyone and specifically to my major because, as a soon to be bio-systems food and processing engineer, it is used to maintain homeostasis.  "In the food industry b uffering solutions in foods play an important role in maintaining specific pH values for optimum activity of enzymes, protein solubility, and functionality. pH may also modify the color and flavor of foods and it is a critical factor in the preservation of many processed foods. Buffering solutions are also used as reaction media in the production of food ingredients and additives. Overall pH control is a major factor ...

The question at hand is? Why is pH so important? How is ph important in our everyday life?  You've come to the right blog.  pH is defined as the measure of hydrogen ion concentration which is used for measuring the acidity or alkalinity of a given solution. Following is the equation that is used for calculating the pH: pH = -log[H + ] It is important for one to assist the body in maintaining a healthy pH because more than often, low levels of pH are linked to diseases and infections. Through the scientific approach we have learned that pH levels are essential to our digestive system.  We know that this includes, the pharynx, stomach, small and large intestines, as well as the rectum and anus. This part of our body creates a pathway that can determine wether you have a healthy life's journey. In the stomach, hydrochloric acid is secreted as food enters the stomach. It turns the pH of the stomach between 1 and 3. This pH is important for the activation of the enzyme pepsin,...

On August 31st we were taught about laboratory safety as well as the use of excel tools/functions to graph certain scenarios within our lab calculations. We simply outlined the safety precautions to be followed as each experiment takes place. The first few rules are the basics of lab safety, washing hands and the use of PPE. We also took in account the procedures to follow in case of an emergency/ accident of hazardous gases and spills. In excel, we were taught how to use the tools included in the program, to produce linear graphs. Along with that we learned how to edit titles, fonts, and slope, etc. . . The specific example we performed in class was an absorbance v. concentration graph. This graph had a linear relationship. The independent variable being the concentration the one that we controlled ( x-axis). This allowed us to practice the functions of excel for the use of our lab reports. Please click the hyperlink "Biochemistry Lab Safety" below to watch an informative vi...