Why is BSA used as a standard for protein content determination? Can other proteins be used instead?
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topic: Preparation of a Calibration Curve
Standard Bovine Serum Albumin (BSA) solution
Why is BSA used as a standard for protein content determination? Can other proteins be used
instead?
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Solved in 2 steps
- 1) Dosage calculation with same units of measure. EX: MD orders 1000mg Ancef. You have available 2000mg in 10mls. How many mls you give?Match the types of data to the example. Precise measurements Assay standard Positive control Quantitative results Negative control Qualitative results Standard curve…Topic: Isolation of Crude Ovalbumin from Egg White by Ammonium Sulfate Precipitation(Salting Out) The computed amount of powdered ammonium sulfate was added to the egg white sample portion byportion with constant stirring while submerged in an ice bath. The solution is expected to become moreturbid, and a white precipitate is expected to form. Explain the purpose of using powdered ammonium sulfate in the isolation of ovalbumin from egg whites.
- topic: Isoelectric Precipitation Appearance of the mixtures containing casein mixed with different buffer solutions: (a) TestTube 1 containing Glycine-HCl buffer; (b) Test Tube 2 containing Acetate buffer; and (c) Test Tube 3containing Phosphate buffer. QUESTION: Based on the given results, what do you think is the isoelectric pH of casein? Briefly discuss your basis for determining it.Bradford Assay Fill in the average A595 and A595 of sample minus A595 of blank. What to do? Calculating the average absorbance of the standards Subtracting the absorbance of the blank (the sample with 0 g/ml) from the absorbance of the other samples Concentration (micrograms/ml) A595 - trial 1 A595 - trial 2 A595 - trial 3 average A595 A595 of sample minus A595 of blank 0 1.501 1.446 1.447 2 1.624 1.558 1.559 5 1.731 1.749 1.712 10 1.901 1.838 1.892 15 2.161 2.108 2.228 18 2.231 2.277 2.319Topic: Reducing and Non Reducing Carbohydrates Among the Benedict's, Reagent, and Iodine tests, which is most highly applicable in biomedical field? Explain
- A series of bovine serum albumin (BSA) was prepared and 1 mL of each solution was subjected to a Bradford assay. The increase in absorbance at 595 nm relative to the concentration of protein was plotted as shown below. Save 1.0- 0.8- 0.6- 0.4- 0.2- 0.04 0.0 0.5 1.0 2.0 2.5 1.5 BSA concentration (mg/mL) a) Using the graph, calculate the original concentration of protein present in a mixture of haemoglobin and methylene blue (diluted 1:50), which gave an absorbance of 0.20 at a wavelength of 595 nm using the Bradford assay. Show all workings. Absorbance (O.D.)Discuss the principles and applications of Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry in molecular diagnostics.Topic: Isolation of Crude Ovalbumin from Egg White by Ammonium Sulfate Precipitation (Salting Out) The computed amount of powdered ammonium sulfate was added to the egg white sample portion byportion with constant stirring while submerged in an ice bath. The solution is expected to become moreturbid, and a white precipitate is expected to form. The resulting mixture was then filtered using a cheesecloth. The residue was discarded, and 30.0 mL of thefiltrate was brought from 40% to 60% saturation by adding the required amount of powdered ammoniumsulfate in the same manner as the previous addition. After adding ammonium sulfate, the mixture was allowed to stand with occasional stirring for 30 minutes inan ice bath. The formation of a white precipitate is expected to happen QUESTION: Explain the significance of allowing the mixture to be submerged in an ice bath for a given time.
- I need to prepare a standard calibration curve for gamma globulin. absorbances on Y and mg of standard protein per assay on X. used 0.1mg/ml gamma stock for tubes 2-6. (Water (ml), gamma (ml), Abosrbance)--> (.036, .004, .290) (.036, .008, .358) (.024, .016, .341) (.016, .024, .520) (.008, .032, .597) - What is the math and how do you get the standard curve?Search for the principles and the uses of the following methods for protein measurements and provide an image that will serve as your visual reference. Serum Protein Electrophoresis Isoelectric Focusing Ion Exchange Chromatography Turbidimetry Solvent FractionationApple puree was analyzed for petulin by HPLC-MS-MS after SPE clean-up. The procedure was 10.0g of puree + 10μl of a 10μg/ml solution of isotopically labeled petulin as internal standard were treated with 10.0ml of pectinase and acetic acid, centrifuged, and filtered. Four ml of the filtrate was passed through a SPE cartridge. The petulin was eluted with 2.0ml of ethyl acetate. The sample was evaporated to dryness and the residue dissolved in 1.0ml of the mobile phase. The analyte signal was 127 and the internal standard 197. Calculate the concentration of petulin in the sample in μg/g (RRF=1).