By Herman S. Overkleeft, Bogdan I. Florea
This quantity makes a speciality of explorative activity-based proteomics,biomedical functions of activity-based proteomics, and chemical thoughts in activity-based proteomics delivering a concise assessment of activity-based protein profiling. Written within the hugely profitable Methods in Molecular Biology series layout, chapters comprise introductions to their respective themes, lists of the required fabrics and reagents, step by step, simply reproducible laboratory protocols, and pointers on troubleshooting and warding off recognized pitfalls.
Authoritative and state of the art, Activity-Based Proteomics: tools and Protocols goals to make sure winning ends up in the additional research of this very important field.
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Extra resources for Activity-Based Proteomics: Methods and Protocols
30. 31. 32. 45 Nat Chem Biol 5(9):616–624. 216 Rappsilber J, Mann M, Ishihama Y (2007) Protocol for micro-purification, enrichment, pre-fractionation and storage of peptides for proteomics using StageTips. Nat Protoc 2(8):1896–1906. 261 White K, Bruckner JV, Guess WL (1973) Toxicological studies of 2-mercaptoethanol. J Pharm Sci 62(2):237–241 Zikolov P, Budevsky O (1973) Acid-base equilibria in ethylene glycol--I: definition of pH and determination of pk-values of acid-base indicators. Talanta 20(5):487–493 Bock VD, Hiemstra H, van Maarseveen JH (2006) Cu-I-catalyzed alkyne-azide “click” cycloadditions from a mechanistic and synthetic perspective.
3 kPa H2O). It evaporates in the pipette (expansion) and presses the liquid out of the pipet. When you pipette ACN, be quick. Take care that you do not drop any ACN on the labels of your Eppendorf tubes. 46. It is beneficial to use a vacuum centrifuge. Alternatively, a desiccator attached to a vacuum pump can also be used to dry your samples although undesirable retardation of boiling may cause your gel pieces to jump out of their Eppendorf tubes. This may lead to a loss of your samples and cause contaminations.
Screw the StageTip holder on a fresh 2 mL tube and place the conditioned StageTip in the hole. Load the peptide solution on the StageTip and centrifuge at 400–800 × g for 2 min (see Note 61). 7. Reapply the flow-through to the StageTip and centrifuge again at 400–800 × g for 2 min (see Note 62). 8. Screw the StageTip holder on a fresh 2 mL tube and place the StageTip with the immobilized peptides in the hole. Screw a clean lid onto the tube containing the flow through; label it properly and save it in case something went wrong with the binding of peptides.
Activity-Based Proteomics: Methods and Protocols by Herman S. Overkleeft, Bogdan I. Florea