Predict the amino acid sequence based on the conditions below. Use the one-letter abbreviation of amino acids.
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- Explain amino acids being amphoteric electrolytes to form peptidebonds. State at least 5 general limitations of Biuret test to detect the existence of peptide bonds in a protein.a. State the importance of using following reagents in SDS-PAGE. 1. Acrylamide 2. Bisacrylamide 3. Tetramethylethylelediamine 4. Glycerol 5. Ammonium persulfate b. Briefly describe the importance of two dimensional electrophoresis in protein separation?Provide examples of amino acids that would yield a positive Azo Dye test and amino acids that would yield a negative Azo Dye test. Show the structures.
- Complete the table below by adding (+) or (-) if eahc sample (amino acid or protein) below will test positive or negative, respectively. sample BIURET NINHYDRIN XANTHOPROTEIC MILLON-NASSE SAKAGUCHI LEAD ACETATE PAULY HOSKIN-COLE G W H Y C R HAIR GELATIN CASEIN PEPTONE Identify the amino acid that will give the following data: BIURET NINHYDRIN XANTHOPROTEIC MILLON-NASSE SAKAGUCHI LEAD ACETATE PAULY HOPSKIN-COLE - + + + - - + - Name of amino acid: structurte of the amino acid:What is the attached enzyme in this assay and its corresponding substrate? A. Horseradish peroxidase and OPD (o-phenylenediamine dihydrochloride) B. Horseradish peroxidase and PNPP (p-Nitrophenyl Phosphate, Disodium Salt) C. Horseradish peroxidase and TMB (3,3',5,5'-tetramethylbenzidine) D. Horseradish peroxidase and ABTS (2,2'-azinobis-3-ethylbenzothiazoleine-6- sulfonic acid)Sketch the appearance after visualization of a protein mixture containing the seven proteins (fibrinogen, y-globulin, collagen, ovalbumin, myoglobin, hemoglobin, insulin) when subjected to two-dimensional (2D) gel electrophoresis.
- Write down a detailed description about the formation of polyacrylamide gel, how it is used for protein separations and the process of electrophoresis in these gels.Give the chemical principles involved in a positive Biuret test, Millon’s Test and Sakaguchi Test. What is the chemical basis of the Ninhydrin Test, Hopkins – Cole test and Sulfur reaction test? What chemical structure should be present in an amino acid or protein to give a positive Xanthoproteic test?You perform a Bradford assay to determine the concentration of isolated α-lactalbumin. You use 50 μL of a two-fold diluted solution of α-lactalbumin in the assay. You generate a standard curve with the following equation for the line: y = 0.163x + 0.082. The absorbance of your sample was 0.674 AU. What is the concentration of α-lactalbumin, in mg/mL, in your sample? Give your answer to three significant figures.
- Discuss the functions and components of Biuret dye in protein content determination. Identify two other methods of protein determination and differentiate them from Biuret method. Why is BSA used as standard for protein content determination? Can other proteins be used instead?Predict the amino acid sequence based on the conditions below. Use the one-letter abbreviation of amino acids. ___1___--_2_- 3 -- --____6_--_7__ a. The amino acid in the C-terminal is optically inactive. b. The fifth amino acid is a neutral aromatic amino acid that yields a negative result for the Xanthoproteic test using intact protein but produces a yellow soln with basic hydrolysate. It also gives a negative result with both Hopkin's Cole and Millon's test using basic hydrolysate. c. After treatment of this peptide with aminopeptidase (specificity: peptide bond on the N-terminus), it yields serine and a hexapeptide d. The amino acid besides the N-terminal amino acid. will yield ornithine and urea after alkaline hydrolysis. e. The third amino acid gives a positive result for Hopkins-Cole Test. f. Chymotrypsin^ cannot cleave the peptide bond on the fifth amino acid because of the presence of this a.a. residue. g. The amino acid at the carboxy-end of the third amino acid will yield a…Consider the following peptide sequence: His-Pro-Lys-Arg-Ala-Gln-Ser-Cys-Pro Which test will the peptide yield a negative result? Group of answer choices Millon's Test Sakaguchi Test Ninhydrin Test Biuret Test