Skip to main content

4. Ion Exchange Chromatography

The tall glass columns which are packed with a synthetic ion-exchange resin is used to carry out the separation. (Resins are the polymers to which various ionizable groups have been chemically added). 



Image result for ion exchange chromatography schematic diagram
Example of anion exchange. 
Resins which have negative charges are called cation exchangers and those which have positive charges are called anion exchangers. When the solution of ions are passed through the column, the ions compete with each other for the charged sites on the resin. The rate of movement of any ion through the column depends on its affinity for the resin sites, its degree of ionization and the nature and concentration of competing ions in the solution. 

Molecules such as proteins and nucleic acids posses a variety of positively and negatively charged groups. For their separations, the differential rates of movement of ions through the column are basis for them. 

Some of anion exchangers are Dowex-2, Bio-Rad AG2, QAESephadex, Diethylam noethylcellulose. 

Some of cation exchangers are caboxymethylkellulose, BioRad AG50, Zeocarb 225. Amberlite IRC50. 

Comments

Popular posts from this blog

Statistics in Flow Cytometry Data and "MFI" values

                                 The speed of the flow cytometry offers wide range of data points and data plots. Due to its sensitivity and versatility, it has been used extensively. Flow cytometer can detect up to 1000s of cells per second.. But, it should be noted that the flow cytometry also involves the statistics, its significance, calculating the fluorescence intensity..                Statistics in flow cytometry involves, total number of data points acquired, percentage population, most importantly mean and median fluorescence intensity, and others. Most of us get confused about the term "MFI", whether it means mean fluorescence intensity or median fluorescence intensity.. Truth is, both.. It means it can be described as either mean or median fluorescence intensity. Some researchers use Mean for MFI and some use Median for MFI values.  (Keep...

Nanodrop

Thermofisher, Nanodrop 1000    Nanodrop is a spectrophotometer which is designed to measure the concentration of nucleic acids at 260nm and Proteins at 280nm present in a very small minute volume. It is also capable of measuring the concentrations of dsDNA, RNA, ssRNA and purified protein. Required sample size is usually 1µL. Nanodrop instruments are usually full spectrum spectrophotometer, i.e., Ultra violet and visible range wavelengths (250nm to 750nm).  Nanodrop is commonly used for the quantification of the DNA concentration and RNA concentration in a given sample. However there are many other applications which can be performed on this instrument.  It works on the principle of Spectrophotometer or Sample Retention System....  Principle of Nanodrop: "The Nanodrop works on the principle of sample retention system.. When the small volume of sample is introduced, the arm and pedestal will work together and make the sample in the form of colum...

Immunophenotyping

Immunophenotyping is a most important and regularly used technique  in Immunology and Immunological research studies. The technique is usually used for the disease monitoring and diagnostic purpose either by immunohistochemistry or by utilizing the high-throughput technique Flow Cytometry.. Flow Cytometry is a common method for the immuno phenotyping the different cell subsets or sub populations of cells. Immunophenotyping is used for the identification and characterisation of the cell surface markers and intracellular markers. In simple terms, the technique is used to detect and study the proteins expressed by the cells. Samples used for the study could be Tissue Section (Fresh or Fixed) or Cell suspension (mono-dispersed cell suspension for FCM). This technique helps in rapid and easy phenotyping of each cell in a heterogeneous sample according to the presence or absence of a protein combination. Markers (or proteins of interest) are usually functional membrane proteins...