Essay on Quantification of Proteins in Solution by...

Experiment 2 Quantification of Proteins in Solution by Spectrophotometer Lab bench# 1 Introduction: Absorption spectroscopy is a common method for finding the concentration of proteins or protein complexes in a solution. Proteins absorb light at specific wavelengths and can be defined by the equation A = log (Io/I). This equation states that an absorbance at a specific wavelength, A is equal to the log of the ratio of incident light intensity (Io), to transmitted light intensity (I). A spectrophotometer can be used quantitatively and qualitatively. A spectrophotometer is used qualitatively to obtain an absorption spectrum, which can be obtained by plotting the absorbance values, over the range of wavelengths tested for the†¦show more content†¦The concentration of protein solution ( µg/ml) calculated using equation of slopes from Fig2 Test Protein | Concentration of Protein Solution ( µg/ml) | | Lowry | Coomassie Blue | UV | BSA | 1.6 | 1.4 | 1.3 | Lysozyme | 1.6 | 1.8 | 5.9 | Hemoglobin | 1.6 | 1.4 | 3.8 | Ovalbumin | 1.6 | 1.9 | 1.5 | Gamma globulin | 1.6 | 2.5 | 2.4 | Discussion: Figure 1 shows the absorption spectrum of stock solution (6x10-5M), p-nitrophenol and 0.02M NaOH, and from the graph it can be inferred that 400nm is the wavelength of maximum absorption because absorption is noted to be the highest at this point.Show MoreRelatedThe Light Energy Absorbed By Colored Compounds1430 Words   |  6 PagespKa of PNP, and to develop standard curves for determination of unknown concentration and pH of unknown samples. Quantification of proteins is needed to determine the progress of protein purification. As the protein becomes more purified, its specific activity will increase as well. In Experiment 4.1, dilutions of PNP are prepared, and the Bradford Method was used to measure protein concentration. There are different types of chromatography, and we would be using gel filtration and affinity. ChromatographyRead MoreThe Light Energy Absorbed By Colored Compounds1429 Words   |  6 PagespKa of PNP, and to develop standard curves for determination of unknown concentration and pH of unknown samples. Quantification of proteins is needed to determine the progress of protein purification. As the protein becomes more purified, its specific activity will increase as well. In Experiment 4.1, dilutions of PNP are prepared, and the Bradford Method was used to measure protein concentration. There are different types of chromatography, and we would be using gel filtration and affinity. ChromatographyRead MoreLab Report On Chemical Tests1279 Words   |  6 Pagessulfate, carbohydrate tests to determine presence of carbohydrates and reducing sugars and, protein test to determine the contaminations present in the fucoidan. 1.5.1 Screening for Carbohydrates 1.5.1.1 Molisch’s Test 2mL of Sulfuric acid will be added to the isolate and 0.2mL alpha napthol. The formation of bluish violet zone indicated the presence of carbohydrates. 1.5.1.2 Fehling’s Test Fehling’s solution (2.5 mL each of Fehling’s AB) will be boiled in a test tube. Equal amount of the isolateRead MoreSpecific Pathogen-Free Animal Rats1525 Words   |  7 Pagesventricular myocardial tissues were collected from each group and fixed in 10% formalin, processed and embedded in paraffin wax. Thin sections of 3-5 microns thickness were cut and placed on microscopic slides. The tissues were deparaffinized in xylene solution, rehydrated in downstream serial dilutions of ethanol and stained with hematoxylin for 10 minutes, bluing for 10 minutes in running tap water, decolorized for 3 seconds by 1% acid alcohol, and the tissues were stained with eosin for 1 minute, washedRead MoreExtraction and Characterization of Proteins3660 Words   |  15 PagesCHARACTERIZATION OF PROTEINS Abstract Different techniques and principles for protein extraction and characterization were demonstrated in this experiment. Various proteins were extracted from different sources: 1.67 g yeast invertase, 1.03 g egg white albumin, and 5.15 g of milk casein. Activity assay for invertase was performed using Benedict’s test and the enzymes inverting action on sucrose was confirmed. Warburg-Christian Method and Bradford Assay were also employed to determine the protein concentrationRead MoreEssay about A Study on A. Nigra Seeds for Pharmaceutical Products1252 Words   |  6 Pagesremove the trace of remaining solvent. The extracts were subjected for vacuum drying in rotary evaporator. The free radical scavenging activity of seed extracts of A. nigra were determined using DPPH whereby solution of DPPH in 99.99% ethanol was prepared and was mixed with ethanolic solutions of n-hexane, ethyl acetate and methanol extracts from A. nigra seeds at determined concentrations. The mixture was shaken and incubated. Butylated hydroxyl toluene (BHT) as positive reference ethanol as negativeRead MoreSex Determination By Amplification Of Amelogenin Gene From Dental Pulp Tissue By Pcr1727 Words   |  7 Pages These are rich sources of DNA and free from contamination by external factors. During the early stages of tooth development, cells of the inner enamel epithelium, differentiate to form ameloblasts synthesizing specific proteins as enamel matrix. Amelogenin being one such protein is present in the dental pulp tissue. The presence of two Amelogenin genes was confirmed by studies of Lau et al. (1989), Shimokawa et al. (1989); one on the X-chromosome and the other on the Y-chromosome. One copy was locatedRead MorePepper Seed Dna Extraction2422 Words   |  10 Pagesgenomic DNA. The sample is lysed in DNAzol (Chomczynski et al. 1997). Both chloroform and ethanol are used to precipitate DNA. These chemicals help separate proteins and polysaccharides from nucleic acids. Because chloroform is denser then water, it will form the bottom layer after centrifuging. This lower layer is what contains the proteins and polysaccharides, leaving the DNA in the upper layer to be collected. DNA is then precipitated and washed with ethanol. TE buffer is used to solubilize theRead M oreA Brief Note On The And Dental Caries And Periodontal Diseases And Non Tobacco Users3209 Words   |  13 Pagesbacteria that acts as an agent in promoting and initializing carious lesions.11 Studies have shown that the bacterium has adhesins which allow it to bind to complementary receptors on a substrate. A. viscosus binds to salivary acidic proline-rich proteins (PRPs) adsorbed onto the apatitic surfaces of the tooth. Cryptic segments exposed in adsorbed molecules are identified by the adhesins, which allow A. viscosus to efficiently attach to teeth while suspended in saliva.12 Actinomyces naeslundii Actinomyces