A Standard Curve Developed From The Cytoplasmic Protein Standards Of Lung Cancer Cells

This Lab Report is an analysis of the results of a two-part experiment. In the first part, we used a gel filtration column to separate the components of a mixture composed of protein and non-protein molecules. By doing so we hoped to obtain fractions that contained single components of the mixture, while also gaining insight into the relative molecular weight of each component compared to each other. We would then plot these fractions onto nitrocellulose paper in order to determine which fractions had protein. In the second part, we would use the fractions which we had determined had protein to conduct an SDS-PAGE. By doing so we hoped to determine an estimate on the molecular weight of the proteins present in each fraction by comparing it to a tracker dye composed of a variety of molecules of differing molecular weight.

In order to examine the variation in sensitivity of lysozyme, gelatin, and BSA, the absorbance reading was obtained using the Bradford and BCA assay

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 through a cuvette filled with Bradford dye and concentrations of PBS and proteins. After the cuvettes are mixed they are placed into a spectrophotometer to measure wavelength. The wavelength given will be used to plot a standard curve based on concentration (x-axis) and wavelength (y-axis). The standard curve is then used to measure an educated guess on the concentrations of unknown protein concentrations. We hypothesized that if we use the Bradford assay and colorimetric spectrophotometry we can determine an accurate concentration of two unknown concentrations of proteins. The results of this lab failed to reject our hypothesis based on accurate measurements of protein concentrations. The standard curves are drawn with a linear increasing slope. The Bradford assay is an accurate way to demine the concentration of an unknown concentration.

A spectrophotometer’s purpose is to use colors of the light spectrum to determine the concentration of light absorbing molecules in a solution. (p.59) In this particular lab, our mission was to determine the protein concentration and the standard curve of the unknown sample of BSA. This, by preparing five dilutions of the unknown solution of BSA together with other known concentrations, and then experimenting by observing how the concentrations were passed through the spectrophotometer. The outcome resolved in the absorption levels being decreased, and this

Cytoplasmic streaming was measured 4 times after flushing the cells with pond water, at 5 minute intervals

Figure 1 contains gel electrophoresis for protein samples. The lanes were labeled from 1 to 10 from the right to the left. Lane 1 contained the ladder fragment. Lane 2 contained the filtrate. Lane 3 contained the S1 sample. Lane 4 contained the P1 sample. Lane 5 contained the P1 medium salt sample. Lane 6 contained the P1 high salt sample. Lane 7 contained the S2 sample. Lane 8 contained the P2 sample. Lane 9 contained the P2 medium salt sample. Lane 10 contained the P2 high salt sample.

Gel electrophoresis method. Qualitative analysis shows protein concentrations in kidney, heart, and liver. 1-6 are kidney tissue. 7-14 are liver and 15-19 are the heart tissue.

The methods of obtaining the cancerous epithelial samples from various areas was a clear way of observing both the level of amplification of CCND1 and the presence of cancer within those tumors or samples. The FISH technique was fitting to localize the specific DNA sequences related to the cause and effect of amplification. Epithelial tissues which contained some development of cancer cells were studied. The researchers used samples that were stored at the University Medical Center Hamburg-Eppendorf, making sure to used samples from areas distant from one another (Burandt, et. al). Taking samples from various areas helped to further see the effect of various amount of amplification of the protein and how it affected the epithelial tissues at different stages.

In order for efficient fixation and processing times to occur, all tissues should be no thicker than 3mm (Washington School of Medicine, 2016). When a tissue sample is thicker than the recommended thickness, penetration and crosslinking of the sample tissue proteins will take an increased amount of time for this to occur. The centre of the tissue may undergo autolysis as the proteins located within the centre of the sample will not be reached at an optimum time. Additionally, the sample may be prevented from undergoing complete fixation by the chosen fixative. However when careful cutting techniques are utilised and the care is taken to cut optimally sized tissue samples, the desired fixation efficiency can be achieved. There are a few processes

Cancer is one of the leading causes of death worldwide as it can develop in almost any organ or tissue. Significant advances in understanding the cellular basis of cancer and the underlying biological mechanisms of tumour has been vastly improved in the recent years (Jiang et al. 1994). Cancer is a genetic disease which requires a series of mutation during mitosis to develop, its characteristics can be associated with their ability to grow and divide abnormal cells uncontrollable while in the mean time invade and cause nearby blood vessels to serve its need. Even though many people are affected by cancer today, the abilities which cancer cells have make it hard to find a single effective treatment for cancer. The focus of research now lies

(Transition: we have discussed the causes, lets look at the risk factors including some environmental ones.)

CFSE solution was added for a final concentration of 0.5M. The mixed packed blood and CFSE solution was then incubated at 37⁰C for 10 mins, then 5 fold volume of PBS was added to it and then the cells were centrifuged for 5 mins at 300 x g. The solution was kept on ice for 5 mins to deactivate, then washed twice with PBS and centrifuged at 300 x g for 5 mins at room temperature. The cells were then resuspended in 10ml DMEM completed medium for cell counting and determining cell viability using trypan blue

Six plastic cups were obtained and labeled 0.2 M, 0.4 M, 0.6 M, 0.8 M, and 1.0 M. Next, each of the six dialysis tubes were knotted on one end and filled with the sucrose samples, and then tied off. These samples were then dried by patting with a paper towel, weighed and placed into their corresponding cups. The mass of each sample was recorded in table 2 on the data sheet. Each cup was then

A1: Triton X-100 is a detergent in laboratory practice and is used to permeabalize unfixed eukaryotic cell membranes and aids in solubilizing membranous proteins. The addition of the substance Triton X-100 would cause the majority of the cell membranes to become permeable thus allowing contents of the eukaryotic cell to leave via simple diffusion hence the large concentration of radioactive prominence to be situated in the supernatant fraction. Again, the detergent property of Triton X-100 is further reinforced by the fact that it causes the majority of all radioactive sulphur to be situated in the supernatant. Without the addition of Triton X-100 the graph displays the greatest quantity of radioactive sulphur to be situated in fraction 3 of which a majority of “small organelles” are situated ranging from a diameter 0.2 to 1.5 μm. It is due to this fact that one can assume the normal cellular location of the CRUSOE1 protein to be situated in normal cellular location in organelles of the previously stated diameter and not it the cytoplasmic structure as the addition of Triton X-100 would assume.

We know that all the incarnate beings are comprised of cells. All these cells found in living organisms have an ability to develop and divide in a precise and controlled way. This ability of cells allows living organisms to function appropriately. Any mutations or changes in these normal functioning cells would lead to an irrepressible growth of these cells. This kind of irrepressible growth is the characteristic feature of cancer cells. Cancer cells are enclosed by a specific membrane, which provides a structural barrier and thus blocks its interaction with microenvironment, receptors, enzymes, and transport systems of human body or living organism. This structural barrier is formed because of the physical and chemical properties of its constituents. Cancer cells also have a property to metastasize. This allows them to spread and localize throughout the. Hence, in order to treat such an unfathomable disease, it is imperative to understand the genesis, cellular structure, and properties of cancerous cells. In this report, I have delineated disparity between healthy and cancer cells, and properties of cancer cells in order to have a better understanding of this unfathomable disease.