📘 ❞ Protein Purification Protocols ❝ كتاب ــ Paul Cutler اصدار 2003
Biology Books - 📖 كتاب ❞ Protein Purification Protocols ❝ ــ Paul Cutler 📖
█ _ Paul Cutler 2003 حصريا كتاب ❞ Protein Purification Protocols ❝ 2024 Protocols: نبذه عن الكتاب: General Strategies Shawn Doonan and Cutler 1 Defining the Problem The chapters that follow in this volume give detailed instructions on how to use the various methods are available for purification of proteins The question arises, however, which these order them achieve any particular case; is, problem must be clearly defined What follows outlines sorts need asked as part definition answers affect approach might taken developing a schedule It should noted here discussion concentrates mainly on laboratory scale isolation Special cases therapeutic at industrial covered Chapters 43 44 (1–5) 1 1 How Much Do I Need? The answer depends purpose protein is required For example, carry out full chemical physical analysis may require several hundreds milligrams purified material, whereas kinetic reaction catalyzed by an enzyme could perhaps done with few less than 1 mg would raise polyclonal antibody At extreme end scale, if the objective obtain limited sequence information from N terminus protein as preliminary design oligonucleotide probe clone screening, then using modern microsequencing techniques, micrograms will sufficient In field of proteomics, previously analytical techniques have become preparative mass spectrometry commonplace sensitive characterization spots gels 36 40–42 describe methodologies These different requirements quantity well dictate source chosen (see Subheading 4 ) will certainly influence large quantities protein requires least early stages, high capacity but low resolving power, such fractional precipitation salt or organic solvents Chapter 13) Process only when content extract has been reduced to manageable levels, medium resolution capacity, ion exchange chromatography 14) can used leading on, if necessary, Biology Books مجاناً PDF اونلاين Biologically Biology natural science concerned study life, its various forms function, organisms interact each other surrounding environment word biology Greek made up two words: bio (βίος) meaning life And loggia ( λογία) means Biology: similarity vegetation animal cover edges African American states, existence same fossil Branches biology Biology ancient thousands years old modern began nineteenth century This multiple branches Among are: Anatomy Botany Biochemia Biogeography Biofisia Cytology cell science Ecology environmental science Development Embryology embryology Genetics genetics Histology histology Anthropology anthropology Microbiology bacteriology Molecular Biology Physiology functions organs Taxonemia taxonomy Virology virology Zoology zoology
كتاب
Protein Purification Protocols
ــ Paul Cutler
صدر 2003م كتاب
Protein Purification Protocols
ــ Paul Cutler
صدر 2003منبذه عن الكتاب:
General Strategies
Shawn Doonan and Paul Cutler
various methods that are available for purification of proteins. The question arises, however, of which of these methods to use and in which order to use them to achieve purification in any particular case; that is, the purification problem must be clearly defined.
What follows outlines the sorts of question that need to be asked as part of that definition and how the answers affect the approach that might be taken to developing a purification schedule. It should be noted here that the discussion concentrates mainly on
laboratory-scale isolation of proteins. Special cases of purification of therapeutic proteins and isolation at industrial scale are covered in Chapters 43 and 44 (1–5).
1.1. How Much Do I Need?
The answer to this question depends on the purpose for which the protein is required.
For example, to carry out a full chemical and physical analysis of a protein may require
several hundreds of milligrams of purified material, whereas a kinetic analysis of the reaction catalyzed by an enzyme could perhaps be done with a few milligrams and less than
1 mg would be required to raise a polyclonal antibody. At the extreme end of the scale, if
the objective is to obtain limited sequence information from the N-terminus of a protein
as a preliminary to the design of an oligonucleotide probe for clone screening, then using
modern microsequencing techniques, a few micrograms will be sufficient. In the field of
proteomics, previously analytical techniques have become preparative with mass spectrometry commonplace for sensitive protein characterization from spots on gels. Chapters 36 and 40–42 describe these methodologies. These different requirements for quantity may well dictate the source of the protein chosen (see Subheading 1.4.) and will
certainly influence the approach to purification. Purification of large quantities of protein
requires use of techniques, at least in the early stages, that have a high capacity but low
resolving power, such as fractional precipitation with salt or organic solvents (see Chapter 13). Process only when the volume and protein content of the extract has been reduced
to manageable levels, methods of medium resolution and capacity, such as ion-exchange
chromatography (see Chapter 14) can be used leading on, if necessary, to high-resolution
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