What is Desalting of peptides?
Desalting is performed by washing with 0.1% TFA in deionized water and peptides are eluted with 80% acetonitrile and 0.1% TFA in deionized water. From: Methods in Cell Biology, 2010.
What is desalting in chromatography?
The method is commonly referred to as desalting when the goal is to remove buffer salts from a sample in exchange for water (with water used to pre-equilibrate the gel-filtration resin). Buffer exchange is the term used when one set of buffer salt in a sample is exchanged for another set.
What is desalting of DNA?
First, no inorganic salts, such as sodium or magnesium, are used when DNA is synthesized chemically. Our standard desalting step refers to the removal of small organic molecules left over from synthesis.
What is desalting in gel filtration?
The method is commonly referred to as desalting when the goal is to remove buffer salts from a sample in exchange for water (with water used to pre-equilibrate the gel-filtration resin). Applications for desalting include: Removing salts from protein solutions.
What is electrostatic desalting?
Desalting is the first refining process applied to crude oil. This is accomplished by washing the oil with water, coalescing small water droplets with electrostatic field forces into drops large enough to settle through the oil to form a bottom water phase and separating the oil and water.
Why dehydration and desalting of crude is necessary?
Electrostatic desalting is generally used, whether employed for oil field production dehydration and desalting or at oil refineries. It is recommended to facilitate the removal of inorganic chlorides and water-soluble contaminants from crude oil.
What is desalting of crude oil?
Desalting involves mixing heated crude oil with washing water, using a mixing valve or static mixers to ensure a proper contact between the crude oil and the water, and then passing it to a separating vessel, where a proper separation between the aqueous and organic phases is achieved.
What is desalting in gel filtration chromatography?
The method is commonly referred to as desalting when the goal is to remove buffer salts from a sample in exchange for water (with water used to pre-equilibrate the gel-filtration resin).
How does a desalter work?
In the desalter, the crude oil is heated and then mixed with 5-15% volume of fresh water so that the water can dilute the dissolved salts. The oil-water mix is put into a settling tank to allow the salt-containing water to separate and be drawn off. Frequently, an electric field is used to encourage water separation.
Is there a no peptide desalting spin column?
No. Protein desalting columns (e.g., Zeba desalting columns) use size exclusion and have a molecular weight cut-off that is typically too high for peptide samples. Peptide desalting, on the other hand, uses reversed-phase chromatography resins (e.g., C18 resin) to bind peptides for salt removal during washing.
Which is the best method for peptide desalting?
A common off-line method of peptide desalting includes the use of columns containing a C18 matrix that binds the digested peptides (stop-and-go extraction tips—STAGE tips) (Rappsilber, Mann, & Ishihama, 2007 ). Ready-made tips are available commercially, but here we propose a custom-made solution:
How many peptides can a spin column bind?
Pierce Peptide Desalting Spin Columns provide a convenient and reproducible way to desalt and remove contaminants from peptide samples prior to analysis by mass spectrometry. Each spin column can bind from 5 µg to 5 mg of native and TMT-labeled peptides.
How much desalting is needed for MS analysis?
Resuspend samples to approximately 1 μg/μL in buffer A for MS analysis. Desalting of peptides is an essential step prior to MS analysis. It improves peptide ionization efficiency and increases the lifetime of the columns.