Can you write areport on ProteinElectrophoresis,Q&A pleaseExperiment No. (7)Title: Protein ElectrophoresisAim of Experiment: Determination of individualprotein in serumTheory:One of the simple techniques for quantitation of serum protein istheir separation into an electric field and this procedure is referred to as"serum protein electrophoresis" (SPE). Electrophoresis is widely used forseparating protein fractions. In addition to separating albumin fromglobulins this technique further fractionates the globulins into four or fivemajor group: alphal (a1), alpha2 (a2) or betal, beta2 (B1 , B2) and gamma(Y) globulins.Electrophoresis is similar in many ways to chromatography, exceptthat separation is due to the movement of charged particles through anelectric current. If these particles are charged differently they will movein opposite directions, the positively charged particles to the cathode andthe negatively charged to the anode. In this separation technique abuffered medium at a fixed pH is essential to ensure that the charge onthe particles and therefore the rate of migration is stabilized.Electrophoresis is especially usedmacromolecular substances such as proteins and lipids, although it isapplicable to many other substances as well. In routine clinical testing thevarious protein substances in human serum are separated and quantitated,enzymes are separated and quantitated, enzymes are separated andidentified, and various hemoglobin fractions are identified. In additionisolation and quantitation of lipoproteins can also be carried out. Theabove substances contain numerous fractions of similar mobility,therefore, when submitted to electrophoresis, the sharpness of separationwill depend upon the extent to which each fraction is homogeneous in itsmigration. Each fraction carries a net charge which varies with the pH ofthe environment.for the separation ofThe direction and speed of electrophoresis depends upon severalfactors such as (1) net charge of the molecule, (2) size and shape of themolecule, (3) electric field strength, (4) nature of the supporting medium,and (5) the temperature of operation. One of the important factors aboveis the pH of the buffer solution. The most commonly used pH for proteinelectrophoresis is pH 8.6, under these conditions most proteins arenegative and therefore migrate toward the anode.

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Can you write a report on Protein Electrophoresis, |Q&A please Aim of Experiment: Determination of individual Experiment No. (7) Title: Protein Electrophoresis protein in serum Theory: One of the simple techniques for quantitation of serum protein is their separation into an electric field and this procedure is referred to as

Can you write a report on Protein Electrophoresis, |Q&A please Aim of Experiment: Determination of individual Experiment No. (7) Title: Protein Electrophoresis protein in serum Theory: One of the simple techniques for quantitation of serum protein is their separation into an electric field and this procedure is referred to as

Principles of Instrumental Analysis

7th Edition

ISBN:9781305577213

Author:Douglas A. Skoog, F. James Holler, Stanley R. Crouch

Publisher:Douglas A. Skoog, F. James Holler, Stanley R. Crouch

Chapter27: Gas Chromatography

Section: Chapter Questions

Problem 27.24QAP

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In order to improve the peptide separation by using a HPLC system, trifluoroacetic acid acts as mobile-phase modifier was added during the preparation of mobile phase. The preparation was performed by a postgraduate student as following:“2.851 g trifluoroacetic acid (MW: 114.02 g/mol) was made up to 500 cm3 in a graduated flask. To this solution, 50 cm3 of ethanol was added, and after mixing the mobile phase was placed in the solvent reservoir and pumping was commenced at 1.5 cm3 min-1.”Based on the given preparation procedure, identify THREE mistakes that were made.
(a) Three components in a mixture have varying distribution constants between the mobile phase and the stationary phase. One of the components has a very high distribution constant. Would this component likely elute first or last and how would you design an experiment or modify the components of the separation to make it perform in the opposite manner? (b) One way to dramatically increase the sensitivity of a quadrupole mass spectrometer is to use a technique called selected ion monitoring (SIM). Explain how this technique works and why it improves sensitivity. Explain what dwell time means. Be sure to draw a diagram and describe how this analysis scheme works. Also, is there a compromise between a wide range of masses scanned and SIM? Explain in easy details.
Protein ABC is a heme-containing protein responsible in electron transfer in microalgae. The target protein was extracted using ammonium sulfate fractional precipitation and quantified using Bradford assay. A set of solutions with increasing concentration was prepared using 15.00 mg/mL analogous protein standard [refer to succeeding table]. Each solution was treated with 80 μL of Bradford reagent and diluted to 1.500mL. The absorbance at 595 nm of each solution was obtained and presented below. Volume of protein std, μL A595 0 0.0002 10 0.0007 25 0.0075 50 0.0165 100 0.0356 250 0.0930 500 0.1950 750 0.3010 1000 0.4175 Report the equation of the calibration curve and proof of linearity (R2 value).
The following data were obtained for four compounds separated on a 20-m capillary column. t, w Compound (min) (min) 8.04 0.15 B 8.26 0.15 8.43 0.16 (a) Explain why it is possible for each compound to have a different number of theoretical plates. (b) calculate the resolution and selectivity factors for each pair of adjacent compounds. The retention time for an unretained solute is 1.19 min.
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Write the differences between physisorption and chemisorption with respect to the following: (i) Specificity (ii) Temperature dependence (iii) Reversibility and (iv) Enthalpy change
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Penicillin F is recovered from a dilute aqueous fermentation broth by extraction with amyl acetate, using 6.5 volumes of solvent per 100. volumes of the aqueous phase. At pH = 3.8 the distribution coefficient KD is 74. (a) What fraction of the penicillin would be recovered in a single ideal stage? (b) What would be the recovery with three-stage extraction using fresh solvent in both stages? (c) How many ideal stages would be needed to give the same recovery as in part (b) if a counterflow cascade were used with V/L = 0.065?
A chromatogram of a mixture of species A, B, and C provided the following data: a) Define "Retention time". b) Calculate the resolution between species A and B. c) Calculate the retention time of species B necessary to separate it from species A with a resolution of 1.5.
Jacqueline isolated an interesting new protein from the broth of a yeast culture. After purification of the protein, she requested an amino acid analysis and found that the protein contained 0.51% tryptophan by weight (tryptophan M, = 204). Calculate the minimum molecular weight of the protein (i.e., assuming there is only one tryptophan residue per protein molecule). minimum M,: Size-exclusion chromatography of the protein gave a molecular weight estimate of 76600. How many tryptophan residues are present in a molecule of the new protein? Enter your answer as a whole number. number of tryptophan residues:
Below are responses generated using the isolated guinea-pig ileum preparation to determine a pA2 value for the cholinergic antagonist homatropine.Responses are expressed in millimetres +/- SDM (n = 4).1. Plot the data on a sheet of paper and then determine by Schild analysis the pA2 value and Schild slope 2. show all calculation on a table form on a sheet of paper 3. discuss the result with conclusion
What are the parameters that can be used for analytes identification and quantification using Cyclic Voltammetry, respectively?