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 recommendations in activity-based proteomics delivering a concise evaluate of activity-based protein profiling. Written within the hugely profitable Methods in Molecular Biology series structure, chapters comprise introductions to their respective themes, lists of the mandatory fabrics and reagents, step by step, effectively reproducible laboratory protocols, and tips about troubleshooting and averting identified pitfalls.
Authoritative and state of the art, Activity-Based Proteomics: equipment and Protocols goals to make sure winning leads to the additional research of this very important field.
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Additional resources for Activity-Based Proteomics: Methods and Protocols
When the solution is homogenous, transfer it to a graduated glass cylinder and adjust the volume to 100 mL with ddH2O. Prepare 10 mL aliquots and store them at −20 °C. The PIC III is added after the labeling reaction to prevent undesirable inhibition of possible targets by the PIC components. 9. g. Bradford assay; use according to manufacturer’s instructions). 26 Susanne Zweerink et al. 10. Dimethyl sulfoxide (DMSO). This is a highly polar organic solvent used for dilution and dissolving of the natural small molecule-based probes.
This makes it hard to adjust the volume in the next step. 3. “Preheated” means that you should not use a cold pipette as this may lead to t-butanol solidification in your pipette. Just rapidly run the pipette a couple of times through a Bunsen burner flame or use an alternative means to slightly warm up your pipette. 4. 0. 0, you should stop adding pellets. Wait until the remaining pellets are dissolved and continue with adding drops of a 1 M NaOH solution. When you take out the pellets from the NaOH bottle, take care.
Nr. 26623). 8. Glycerol is a viscous liquid that can be hard to handle. To dispense this liquid, always calculate the required weight (using the density of glycerol), place your tube on a balance and directly pour in the calculated amount. 26 g·cm−3) directly to your Falcon tube. 9. β-Mercaptoethanol is a strong reducing agent. It is toxic  and smells bad. Wear a lab coat, gloves and protective glasses at all times when handling this chemical or samples containing it. Always work in a chemical hood especially when boiling samples containing β-mercaptoethanol.