CryoBioPhysica.com, pISep Advanced Protein Purification Technology

click on image for detailed view

Under Development, Coming Soon from CryBioPhysica. Inc.
pISep pH Gradient Maker Plus
pISep Advanced Chromatographic Software Applications

NaCl Switch
With this tool it is simple to add any background concentration of NaCl, 0-1 M, to your pISep buffers. Input that NaCl concentration into pISep pH Gradient Maker Plus and construct your pH gradient just as you would in pISep pH Gradient Maker, press the PLOT button and instantly get the values of %B (and %A) vs. column volumes that you need to program your FPLC or HPLC in order to create your pH gradient at the desired background concentration of NaCl.
Urea Switch
It is also very simple to add any background concentration of urea, 0-8 M, to your pISep buffers. Input that urea concentration into pISep pH Gradient Maker Plus and construct your pH gradient just as you would in pISep pH Gradient Maker, press the PLOT button and instantly get the values of %B (and %A) vs. column volumes that you need to program your FPLC or HPLC in order to create your pH gradient at the desired urea concentration.
Acetonitrile Switch
If all the above isn’t enough you can add any background concentration of acetonitrile, 0-80%, to your pISep buffers. Input the acetonitrile concentration into pISep pH Gradient Maker Plus and construct your pH gradient just as you would in pISep pH Gradient Maker, press the PLOT button and instantly get the values of %B (and %A) vs. column volumes that you need to program your FPLC or HPLC in order to create your pH gradient at the desired acetonitrile concentration.

What are the benefits and what I can do?

You can use the NaCl chromatographic tool to separate proteins at more neutral pH values by shifting the pH at which the target protein exits the column, its apparent pI. Since each protein shifts its apparent pI as a function of salt concentration and pH gradient structure, this tool greatly enhances isoform resolution capability.
You can use these new, unprecedented urea and acetonitrile chromatographic tools to separate transformationally expressed proteins trapped in insoluble inclusion bodies better and faster than with size exclusion chromatography.
You can solubilize and fractionate complex mixtures of membrane proteins.
You can unmask a hidden relationship between apparent pI and the configurational state of your protein. For instance, proteins differing by one or two nonpolar amino acids won’t normally show any difference in apparent pI, but if they differ in stability their differential response to urea or acetonitrile will allow pISep to separate them. Flowing the eluant from an IEX column running a pISep gradient with added acetonitrile directly into an RP column will also provide a flexible new method of separating difficult to purify proteins.
You will get an exciting new tool for studying protein folding/unfolding. Long lived multiple, dynamically stable partially unfolded forms of a pure protein can be easily created and separated using pISep and urea or acetonitrile.
With this three switch system you get everything you need for utilizing the most versatile pH gradient system on the market.

	click on image for detailed view
Under Development, Coming Soon from CryBioPhysica. Inc. pISep pH Gradient Maker Plus pISep Advanced Chromatographic Software Applications NaCl Switch With this tool it is simple to add any background concentration of NaCl, 0-1 M, to your pISep buffers. Input that NaCl concentration into pISep pH Gradient Maker Plus and construct your pH gradient just as you would in pISep pH Gradient Maker, press the PLOT button and instantly get the values of %B (and %A) vs. column volumes that you need to program your FPLC or HPLC in order to create your pH gradient at the desired background concentration of NaCl. Urea Switch It is also very simple to add any background concentration of urea, 0-8 M, to your pISep buffers. Input that urea concentration into pISep pH Gradient Maker Plus and construct your pH gradient just as you would in pISep pH Gradient Maker, press the PLOT button and instantly get the values of %B (and %A) vs. column volumes that you need to program your FPLC or HPLC in order to create your pH gradient at the desired urea concentration. Acetonitrile Switch If all the above isn’t enough you can add any background concentration of acetonitrile, 0-80%, to your pISep buffers. Input the acetonitrile concentration into pISep pH Gradient Maker Plus and construct your pH gradient just as you would in pISep pH Gradient Maker, press the PLOT button and instantly get the values of %B (and %A) vs. column volumes that you need to program your FPLC or HPLC in order to create your pH gradient at the desired acetonitrile concentration. What are the benefits and what I can do? You can use the NaCl chromatographic tool to separate proteins at more neutral pH values by shifting the pH at which the target protein exits the column, its apparent pI. Since each protein shifts its apparent pI as a function of salt concentration and pH gradient structure, this tool greatly enhances isoform resolution capability. You can use these new, unprecedented urea and acetonitrile chromatographic tools to separate transformationally expressed proteins trapped in insoluble inclusion bodies better and faster than with size exclusion chromatography. You can solubilize and fractionate complex mixtures of membrane proteins. You can unmask a hidden relationship between apparent pI and the configurational state of your protein. For instance, proteins differing by one or two nonpolar amino acids won’t normally show any difference in apparent pI, but if they differ in stability their differential response to urea or acetonitrile will allow pISep to separate them. Flowing the eluant from an IEX column running a pISep gradient with added acetonitrile directly into an RP column will also provide a flexible new method of separating difficult to purify proteins. You will get an exciting new tool for studying protein folding/unfolding. Long lived multiple, dynamically stable partially unfolded forms of a pure protein can be easily created and separated using pISep and urea or acetonitrile. With this three switch system you get everything you need for utilizing the most versatile pH gradient system on the market.
Copyright © 2005, CryoBioPhysica, Inc.