(15) Identify the best technique for purifying or characterizing proteins. [In t
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Question
(15) Identify the best technique for purifying or characterizing proteins. [In the space beside each property (left), write a letter (A- E) corresponding to the best technique (right).] Protein Property separation by solubility in water separation by mass separation by positive charge three-dimensional atomic structure location within a developing embryo Biochemical Techniqu A) gel electrophoresis (using SDS) B) electron diffraction (using X-ray) C) precipitation (using NHSO) D) chromatography (using CM-cellulose resin) E) microscopy (using fluorescent antibodies)Explanation / Answer
the correct match is given below
Separation by solubility in water – precipitation (using NH4SO4)
NH4SO4 is used to separate proteins by altering their solubility. It increses the solubility of proteins at low concentrations (salting in) and decreases the protein solubility at high concentrations (salting out).
Separation by mass – gel electrophoresis (using SDS)
In gel electrophoresis, the protein molecules are separate on the basis of their masses by applying electric field. SDS is uses to to provide the net negative charge to protein molecules so that they can move towards the positively charged electrode, while the pore size of agarose provides mass based separation.
Separation by positive charge – chromatography (using Cm-cellulose resin)
Ion exchange chromatography constitutes the separation of ions and polar molecules based on their affinity to the ion exchanger. CM Cellulose resin is designed for the cation exchange chromatography i.e. it is involved in the separation of positively charged molecules.
Three dimentional atomic structure – electron differaction (using X-ray)
Location within a developing embryo – microscopy (using fluorescent antibodies)
Flourescent antibodies bind to the target protein and colocalize with it. This localization can be easily determinent by flourescent microscope.
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