Gel Filtration
Gel filtration
GEL FILTRATION OF PROTIENS
Aim:
The aim of this experiment is to identify proteins from a complex mixture using the gel filtration technique also known as size exclusion chromatography. This technique is widely used by biochemists when proteins larger than the pores are excluded from the column and the smaller molecules elute last and then collected in test tubes for examination by spectroscopic techniques.
The red/brown proteins, in particular, will be observed closely as they can either by haemoglobin or cytochrome c, using the spectrometer their absorbance readings will determine if they are haemoglobin or cytochrome c.
Background research:
A technique widely used by scientists for over 30 years gel filtration chromatography purifies biological samples according to their molecular size and weight. The procedure involves a column of beads, containing within various pores which allows the heaviest molecule to penetrate through the column and eluting first, and then decreasing in size and weight in order. Globular proteins have different rates of migration due to their size and weight the heaviest proteins cannot fit to the channels of the interior beads (pores) therefore it cannot pass through and migrate quickly down the column, whereas the smaller molecule that has the same size diameter with that of the pores will migrate through the pores and will take longer as they are less in weight and they will take the longest because the pores that match in diameter will have to be pass through and have to travel in distance more than the heavier molecules
Gel filtration is an easy technique which enables biochemists to analyse many biological samples to estimate molecular weights and for purification. For example globular proteins, urea, phenol, enzymes, antigens, DNA. The proteins will be collected into the fractions and then examined by a spectrometer for identifying which globular protein the fraction consists
GEL FILTRATION OF PROTIENS
Aim:
The aim of this experiment is to identify proteins from a complex mixture using the gel filtration technique also known as size exclusion chromatography. This technique is widely used by biochemists when proteins larger than the pores are excluded from the column and the smaller molecules elute last and then collected in test tubes for examination by spectroscopic techniques.
The red/brown proteins, in particular, will be observed closely as they can either by haemoglobin or cytochrome c, using the spectrometer their absorbance readings will determine if they are haemoglobin or cytochrome c.
Background research:
A technique widely used by scientists for over 30 years gel filtration chromatography purifies biological samples according to their molecular size and weight. The procedure involves a column of beads, containing within various pores which allows the heaviest molecule to penetrate through the column and eluting first, and then decreasing in size and weight in order. Globular proteins have different rates of migration due to their size and weight the heaviest proteins cannot fit to the channels of the interior beads (pores) therefore it cannot pass through and migrate quickly down the column, whereas the smaller molecule that has the same size diameter with that of the pores will migrate through the pores and will take longer as they are less in weight and they will take the longest because the pores that match in diameter will have to be pass through and have to travel in distance more than the heavier molecules
Gel filtration is an easy technique which enables biochemists to analyse many biological samples to estimate molecular weights and for purification. For example globular proteins, urea, phenol, enzymes, antigens, DNA. The proteins will be collected into the fractions and then examined by a spectrometer for identifying which globular protein the fraction consists