About the Author - Peter Nollert

Peter Nollert

I'm Peter Nollert and I write this blog to point researchers to topics that are relevant to protein crystallization. My mission is to help spread knowledge that is 'out there on the web' and help you succeed with your protein structure research.  I oversee the membrane protein research and technology development activities at Emerald BioStructures. Check out The GPCR blog, or my publications

Blog Archive

Protein Crystallization Hits

Covering protein crystallization space Part II: which Buffers and Salts do you really need?

by Peter Nollert
April 17, 2012 23:47

This is the continuation of a target agnostic survey of often used protein crystallization reagents, based on data obtained from the Biological Macromolecule Crystallization Database (BMCD ver. 4.03). The question I'm trying to address is: which buffers and salts should you inventory?  

Covering protein crystallization space with PEGs seemed a simple affair: a set of only 12 different Polyethyleneglycols is sufficient to formulate ca. 88% of all PEG-based protein crystallization conditions.  

 

The situation is much less clear cut for buffers and salts that are relevant to protein crystallization. Shy of half of all protein crystallizations listed in the (BMCD ver. 4.03), 45%, can be carried out with 8 different buffers (see Fig. below). Tris buffer seems to be the champion. I interpret this as a result of investigator bias rather than there being a solid scientific reason for this buffer to play such an important role. My explanation is that neutral pH Tris buffers dominate the lab bench, and researchers take what they find first… If this is in fact true, it could support the notion that the nature of the buffer is of somewhat low importance for many protein crystallizations.

 

Salts are much more interesting since they  can have a dramatic effect on water properties and protein surface decoration, both affecting the ordered association of protein molecules into a crystal. Ammonium sulfate, the classic protein precipitation reagent is the clear winner. Curiously, several of the salts, such as citrates, phosphates and acetates, - the ones that provide both high ion strength and pH buffer capacity are fairly high ranked.

 

Popular protein crystallization buffers and salts as extracted from the Biological Macromolecule Crystallization Database (BMCD ver. 4.03)

 

When it comes to warehousing stock solutions for simple and quick preparation of optimization screens, these buffers: CHES, CAPS, Bicine, Tris, Hepes, Imidazole, Bis-Tris, MES

and these salts: magnesium acetate, lithium nitrate, calcium chloride, zinc acetate, potassium/sodium tartrate, sodium citrate, sodium chloride, sodium phosphate, potassium citrate, magnesium sulfate, lithium chloride, calcium acetate, ammonium phosphate, ammonium sulfate are a good start. 

Tags: Best practice | Optimization | Product Information | Protein Crystallization

Feeding Your Robot: Taking Chaos Out of Your Wet Lab

by Peter Nollert
April 3, 2012 23:30

Join Us For a Free Webinar April 11th "Feeding Your Robot: Taking Chaos Out of Your Wet Lab"

Date: April 11, 2012

Time: 11:00 AM - 11:50 PM PDT

Speakers: Peter Nollert, Ph.D., Chief Technologist and Amy Syverson, Consumables Sales and Support Manager

Register

Liquid handling instrumentation in life science laboratories can dramatically increase the quantity and quality of research output. On the flip side, these 'robots' require a supply of reagents and buffers to perform. And when your robots are not properly 'fed', research output can be severely hampered. In this webinar you will learn 5 best practices to avoid chaos in the wet lab. Our advice is based on years of experience in feeding our own liquid handling instrumentation with stock solutions for the production of protein crystallization reagent kits. Webinar attendees will learn the fundamentals of proper stock solution management and how to apply it to their own research, making liquid handling instrumentation a successful component in the lab.

Be more productive, register for our April 11th webinar today.

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