Answer the following study questions in your laboratory notebook or on a 1. What

Question

Answer the following study questions in your laboratory notebook or on a 1. What is the basis for the separation of different compounds by ion ex- 2. How can molecules with the same charge at varying amounts be sepa- 3. Why are celluloses often used as supports to separate large biologically 4. What do you think would happen if 0.5M K' acetate were used first to Why is it important to prepare a standard curve for each spectropho- separate worksheet change? rated by chromatography? active proteins? elute the sample? Why? 5. tometer?

Explanation / Answer

Ans 1) The basis for the separation of different compounds by ion exchange is the presence of charge present on the proteins or the nucleotides. The positive and negative group or side chain brings the difference. The net charge of the compound like protein, amino acid, nucleic acid or peptide helps in brings a separation.

Ans 2) The molecules with same charge at various amounts be separated by chromatography as it moves along the chromatography according to the size. The separation can be done based on size where the large ones tend to move at greater speed as compared to the smaller ones.

Ans 3) The celluloses often used as support to separate large biologically active proteins as it imparts a stronger interaction. With more stronger interaction of the protein with DNA, the protein can be eluted in presence of higher salt concentration. In case of the cellulose, there is some limit to the substitution number that can be made per unit of cellulose.

Ans 4) 0.5M K+ acetate were used first to elute the sample because it has more stronger attraction force which selectively attracts the molecules for the process of elution.

Ans 5) It is important to prepare the standard curve for each spectrophotometer as it is used in quantitative research technique. It helps in determining the properties of the unknown samples by interpolation on the graph. It helps in measuring and determining the capacity of a substance to absorb light of specified wavelength.

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