By Robert L. Switzer
This variation of Experimental Biochemistry is an ideal healthy for today’s biochemistry laboratory course. It offers complete insurance of significant options utilized in modern biochemical examine and offers scholars the history thought they should comprehend the character of the experiments. completely school room verified, the experiments comprise the entire variety of biochemical fabrics in an try to simulate paintings in a learn lab. additionally, a complete appendix presents particular approaches for training of reagents and fabrics, in addition to priceless feedback for the instructor. Custom Publishing Create a personalised model of this article or mix'n'match it with comparable titles with W.H. Freeman customized Publishing!
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Additional resources for Experimental Biochemistry
5 pH units away from the operating pH of the column. Remember that the charged groups on the ion-exchange resin will electrostatically attract any molecule or ion carrying an opposite charge. Therefore, it is preferable to use a buffer whose charged form is of the same sign as that on the resin. For example, Tris is a buffer commonly used in conjunction with anionexchange resins. Tris can either exist in the protonated form (positively charged) or the unprotonated (neutral) form (ClϪ is the counter-ion for Tris in the protonated form).
Affinity chromatography is perhaps the most rapidly advancing type of chromatography in the field of biochemistry. Considering that the goal of any chromatographic step is to achieve the highest yield and the highest degree of purity possible, affinity chromatography presents itself as the most promising candidate for this purpose. If one considers a biological molecule such as an enzyme, what distinguishes one enzyme from the hundreds that are found in the cell? ) for which the enzyme has affinity.
The behavior of a compound in thin-layer partition chromatography is usually described in terms of relative mobility (Rf ): Distance traveled by sample (cm) Rf ϭ ᎏᎏᎏᎏᎏ Distance traveled by solvent front (cm) The larger the value of Rf, the more affinity the compound has for a particular mobile phase solvent. The major advantages of thin-layer partition chromatography are cost and versatility. Compared to column chromatography and HPLC, thin-layer chromatography is very inexpensive to perform. Experimentation with different adsorbents and mobile-phase compositions will allow the separa- 39 40 SECTION I Basic Techniques of Experimental Biochemistry tion of a wide variety of biological molecules.