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Thanks to those who replied. The humour comes of course from a slight feeling that there is some truth in the 'respectful' version. Proteins are wonderful subtle machines so perhaps we should be careful and thoughtful when we expose them to the horrors of most crystallization conditions! Which takes me to the original point of my search - which was for a dialysis method for lysozyme. I was trying to set up a micro dialysis format for my protein but wanted to test it first. I am using spectrapore 8K dialysis membrane and wondered if that would retain lysozyme enough to get crystals (I'd like to stick with this cut-off as the physical and sealing properties of other membranes might be different). Or if someone has a higher MW cheap alternative protein/condition? Best wishes - agus 'Bliadhna Mhath Ur Dhuibh Uile' as we say in gaelic. Martyn ----- Martyn Symmons Cambridge ----- Original Message ---- From: David Briggs <[log in to unmask]> To: [log in to unmask] Sent: Monday, 21 December, 2009 17:26:15 Subject: Re: [ccp4bb] where I have been going wrong in crystallization? Hi Martyn, this recipe tends to work for me... Lysozyme: 50 mg/ml in 0.1 M Sodium Acetate pH 4.8 Reagent: 8% w/v Sodium Chloride, 0.1 M Sodium Acetate pH 4.8 Additional Reagents: Index Reagent 8, 22, 28, 31, 34, 40, 58, 59, 69, 86, 88 Mix equal amounts of lysozyme with reagent, incubate at 4 or 22 degrees Celsius. Batch or vapor diffusion works fine. ==> Direct copy/paste from http://hamptonresearch.com/experiments.aspx <== HTH and Happy Christmas. Cheers, Dave ============================ David C. Briggs PhD ============================ University of Manchester E-mail: [log in to unmask] ============================ Twitter: @xtaldave Skype: DocDCB ============================ 2009/12/21 MARTYN SYMMONS <[log in to unmask]>: > Dear All > checking out the Lysozyme crystallization methods on the web I liked the Rigaku Instructions that I found: > (http://www.rigaku.com/protein/crystallization.html) > > "...create a drop of 3ul lysozyme solution, and 3 ul of well solution, respectfully, for a total drop size of 6ul..." > > So perhaps sometimes I am just not respectful enough to deserve crystals ? > > good wishes to all > regards, > Martyn > ----------------------- > Martyn Symmons > MRC-MBU Cambridge UK > 'Chan fhiosrach mur feòraich.' > Gaelic proverb - > Nothing asked, nothing learned. >