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This programme is one of a series of six which concentrate on crucial biochemical techniques, then practice and applications, presented in a studio-based laboratory setting. After a brief introduct...ion by Professor Rose, Dr. Brian Tiplady demonstrates the principles and practice of polyacrylamide-gel electrophoresis. He demonstrates how this technique shows up the differences between the samples obtained in the purification of yeast hexokinase (see Programmes 1 and 2). Jeff Haywood demonstrates how the combination of electrophoresis and chromatography on paper can be used to show up the chemical differences between similar proteins. This technique is known as protein 'fingerprinting'. He uses as his examples haemoglobins - from normal human adults, from adults suffering from sickle cell disease, and from babies. Finally Dr. Tiplady uses a spectrophotometer to show how it is possible to monitor the uptake of oxygen by human blood. From such experiments it is possible to show that the previously demonstrated structural difference between the haemoglobin of a baby and its mothers is of functional importance while the baby is in the womb.
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Description This programme is one of a series of six which concentrate on crucial biochemical techniques, then practice and applications, presented in a studio-based laboratory setting. After a brief introduct...ion by Professor Rose, Dr. Brian Tiplady demonstrates the principles and practice of polyacrylamide-gel electrophoresis. He demonstrates how this technique shows up the differences between the samples obtained in the purification of yeast hexokinase (see Programmes 1 and 2). Jeff Haywood demonstrates how the combination of electrophoresis and chromatography on paper can be used to show up the chemical differences between similar proteins. This technique is known as protein 'fingerprinting'. He uses as his examples haemoglobins - from normal human adults, from adults suffering from sickle cell disease, and from babies. Finally Dr. Tiplady uses a spectrophotometer to show how it is possible to monitor the uptake of oxygen by human blood. From such experiments it is possible to show that the previously demonstrated structural difference between the haemoglobin of a baby and its mothers is of functional importance while the baby is in the womb.
| Item code: | S2-1; 03 |
|---|---|
| First transmission date: | 22-07-1972 |
| Published: | 1972 |
| Rights Statement: | Rights owned or controlled by The Open University |
| Restrictions on use: | This material can be used in accordance with The Open University conditions of use. A link to the conditions can be found at the bottom of all OUDA web pages. |
| Duration: | 00:24:10 |
| + Show more... | |
| Producer: | Roger Jones |
| Contributors: | Stephen Rose; Brian Tiplady |
| Publisher: | BBC Open University |
| Keyword(s): | Applications; Biochemical techniques; Blood oxygen; Haemoglobin; Polyacrylamide-gel electrophoresis; Practice; Protein fingerprinting; Spectrophotometer |
| Footage description: | Stephen Rose introduces the programme. It will examine the technique of polyacrylamide gel electrophoresis as a criterion for determining sample purity, and determining the difference in structure and function between very similar proteins. Brian Tiplady with a gel containing the starting material for the yeast hexokinase purification. The proteins have been separated into distinct layers by passing an electric current through the gel. These are seen against back lighting. Tiplady explains and demonstrates the technique for obtaining this separation in a gel. Tiplady stains the sample so that the protein layers become visible. Tiplady with 3 tubes containing samples from stages in the purification of hexokinase. These have been separated in gels by the electrophoresis process. Tiplady compares their respective bands of protein. Tiplady with 3 tubes containing hemoglobin. He introduces Jeff Haywood who will examine them Jeff Haywood with the 3 Hemoglobin samples in a gel. The samples are: 1) normal adult hemoglobin, 2) adult hemoglobin from sickle cell anemia sufferer, 3) Normal adult plus that of a newborn baby hemoglobin. Haywood explains that to analyse the differences requires two stages - 1) paper chromatography. 2) high voltage electrophoresis. These processes are known as "finger printing" of proteins. Haywood demonstrates the technique of paper chromatography. He shows the result. Haywood next demonstrates and explains the technique of high voltage electrophoresis which further separates the proteins in the sample. He shows the results. Haywood compares the results of the three samples (normal, fetal, and sickle cell hemoglobin). Brian Tiplady explains and demonstrates how to plot the oxygen uptake curves for the hemoglobins. He uses a spectrophotometer to take his readings which are monitored on a pen recorder graph. Tiplady compares the oxygen uptake curves for two of the samples (fetal and normal adult) and relates them to the functional properties of the organism. Stephen Rose sums up the programme. |
| Master spool number: | 6LT/70438 |
| Production number: | 00521_2129 |
| Videofinder number: | 2817 |
| Available to public: | no |
