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Description
This programme is one of a series of six which concentrate on crucial biochemical techniques, their practice and applications, presented in a studio-based laboratory setting. In this programme the ...sixth (dialysis) stage of a seven stage purification of the enzyme yeast hexokinase is completed. After dialysis the last, seventh stage of column chrometography on DEAE cellulose is demonstrated. The solutions to the problems of monitoring and collecting proteins as they are separated on the column are demonstrated, using ultraviolet spectrophotometry and a fraction collector. The results of the purification achieved are discussed and the question of the degree of purity is raised.
This programme is one of a series of six which concentrate on crucial biochemical techniques, their practice and applications, presented in a studio-based laboratory setting. In this programme the ...sixth (dialysis) stage of a seven stage purification of the enzyme yeast hexokinase is completed. After dialysis the last, seventh stage of column chrometography on DEAE cellulose is demonstrated. The solutions to the problems of monitoring and collecting proteins as they are separated on the column are demonstrated, using ultraviolet spectrophotometry and a fraction collector. The results of the purification achieved are discussed and the question of the degree of purity is raised.
| Item code: | S2-1; 02 |
|---|---|
| First transmission date: | 08-07-1972 |
| Published: | 1972 |
| Rights Statement: | |
| Restrictions on use: | |
| Duration: | 00:24:28 |
| + Show more... | |
| Producer: | Roger Jones |
| Contributors: | Norman Cohen; Stephen Rose |
| Publisher: | BBC Open University |
| Keyword(s): | Applications; Biochemical techniques; DEAE cellulose chrometography; Degree of purity; Enzyme purification; Fraction collector; Practice; Stage 6-7; Ultraviolet spectrophotometry |
| Footage description: | Steven Rose introduces the programme. He sums up the steps taken so far (in unit 1) in the purification of hexokinase from yeast. Rose carries on where the previous programme left off - the dialysis of amonium sulphate from the solution. Rose removes the solution from dialysis and empties it into a test tube. The solution is a partially purified Hexokinase. N.R.Cohen explains how the electric charge often present in proteins can be used to further purify the hexokinase. This introduces the technique of chromatography. Cohen with the apparatus for ion chrometography. He explains how it works. Cohen demonstrates the apparatus, particularly the part which automatically measures samples of pre-determined size into test tubes. Cohen with the utltra-violet spectrophotometer. He explains how it works. Steven Rose with the pen recording from the spectrophotometer. He explains that the peaks in the recording represent proteins from the column in the ion chromatography purification. Rose assays samples corresponding to the peaks on the pen recording. The test is based on colour change in the ph activity of the hexokinase. |
| Master spool number: | 6LT/70323 |
| Production number: | 00521_2128 |
| Videofinder number: | 2816 |
| Available to public: | no |
