Definition of Gel Electrophoresis: Gel Electrophoresis is the process in which molecules (such as proteins, DNA, or RNA fragments) can

be separated according to size and electrical charge b appl ing an electric current to them! "he current forces the molecules through pores in a thin la er of gel, a #ell $li%e substance! "he gel can be made so that its pores are #ust the right dimensions for separating molecules within a specific range of sizes and shapes! &maller fragments usuall tra'el further than large ones! "he process is sometimes called gel electrophoresis! Gel Electrophoresis is electrophoresis performed in silica gel, a porous inert medium! Electrophoresis is a process in which colloidal particles or macromolecules (nucleic acids or proteins) with a net electric charge migrate in a solution under the influence of an electric current (also %nown as cataphoresis or %ataphoresis)! Electro refers to the energ of electricit , and phoresis deri'es from the Gree% 'erb phoros, meaning (to carr across)! &eparation of macromolecules (or large molecules) depends upon two forces, namel mass and charge!

Principles of Gel Electrophoresis

Electrophoresis is a technique used to separate and sometimes purify macromolecules - especially proteins and nucleic acids - that differ in size, charge or conformation. As such, it is one of the most widel $ used techni*ues in biochemistr and molecular biolog !

When charged molecules are placed in an electric field, they migrate toward either the positive or negative pole according to their chargeProteins and nucleic acids are electrophoresed within a "gel". +ost commonl , the gel is cast in the shape of a thin slab, with wells for loading the sample! "he gel is immersed within an electrophoresis buffer that pro'ides ions to carr a current and some t pe of buffer to maintain the p, at a relati'el constant 'alue! he gel itself is composed of either agarose or polyacrylamide, each of which ha'e attributes suitable to particular tas%s: !garose is a pol saccharide e-tracted from seaweed! .t is t picall used at concentrations of /!0 to 12! "he higher the agarose concentration the 3stiffer3 the gel! Agarose gels are e-tremel eas to prepare: ou simpl mi- agarose powder with buffer solution, melt it b heating, and pour the gel! .t is also non$to-ic! !garose gels have a large range of separation, "ut relatively low resolving power. 4 'ar ing the concentration of agarose, fragments of DNA from about 1// to 0/,/// bp can be separated using standard electrophoretic techni*ues! Polyacrylamide is a cross$lin%ed pol mer of acr lamide! "he length of the pol mer chains is dictated b the concentration of acr lamide used, which is t picall between 5!0 and 1/2! 6ol acr lamide gels are significantl more anno ing to prepare than agarose gels! 4ecause o- gen inhibits the pol merization process, the must be poured between glass plates (or c linders)! Polyacrylamide gels have a rather small range of separation, "ut very high resolving power.

"76E& 89 E:E;"R86,8RE&.& #ne-$imensional %.&odium dodecyl sulphate polyacrylamide gel electrophoresis '&$&-P!GE( &D&$6AGE is a common form of electrophoresis for anal zing proteins! "he &D& part of the name is a protein denaturing detergent that causes the molecule to unfold! "he protein pol peptides mo'e through the gel at different rates depending on mass, allowing researchers to stud proteins based on size! ).*soelectric +ocusing'*E+( .n .soelectric focusing, proteins are separated b electrophoresis in a p, gradient based on their isoelectric point(pl)! At all p,s other than their isoelectric point, proteins will be charged! .f the are positi'el charged, the will mo'e towards the more negati'e end of the gel and if the are negati'el charged the will mo'e towards the more positi'e end of the gel! At its isoelectric point, since the protein molecule carr no net charge it accumulates or focuses into a sharp band! ,.-ative-P!GE Nati'e 6AGE is used to separate proteins in their nati'e states according to difference in their charge densit ! .t is used for prepartion of purified and acti'e proteins! .n nati'e 6AGE the mobilit depends on both the protein<s charge and its h drod namic size! Nati'e 6AGE can be carried out near neutral p, to a'oid acid or al%aline !"he apparatus is %ept cool to minimize denaturation of proteins and proteol sis! "wo Dimensional 6ol acr lamide gel Electrophoresis(1D$6AGE)

1D Electrophoresis is a techni*ue efficient separation techni*ue for proteins! "his techni*ue results in gels that contain spots! Each spot on the gel corresponds to a different protein! "hen the gels are stained similarl as to =D anal sis! 1D Electrophoresis is widel used, and certain methods of it can be coupled with mass spectrometr in order to identif proteins! Differential in Gel Electrophoresis(D.GE) Differential in Gel Electrophoresis (D.GE) is a techni*ue to monitor the differences in proteomic profile between cells in different functional states! "his technolog allows for simultaneous separation and comparison of up to three samples on one gel >6N;$6AGE
>uantitati'e 6reparati'e Nati'e ;ontinuous 6ol Acr lamide GelElectrophoresis is a techni*ue to isolate acti'e or nati'e metalloproteins (e!g!, metal chaperones, prions, metal transport proteins, am loids, metalloenz mes, metallopeptides) in biological samples and to resol'e properl and improperl folded metal cofactor$containing proteins in compleprotein mi-tures

;apillar ;apillar electrophoresis is a method similar to &D&$6AGE! .t separates molecules based on their charge and mass! +olecules are placed in rows called capillaries filled with conducti'e, electrol te fluid! "his method is an older techni*ue introduced in the =?@/s! !pplications of Electrophoresis

%. $-! !nalysis
Electrophoresis is one wa of anal zing DNA, which is the code that contains all the traits ou inherited from our parents! DNA is arranged in se*uences,

one se*uence represents the color of our e es and another se*uence represents the color of our s%in! "hrough electrophoresis, specific DNA se*uences can be anal zed, isolated and cloned

). !nti"iotics !nalysis
"he application of electrophoresis in antibiotic studies dates bac% to the =?0/s! 9urther studies led to impro'ed electrophoretic techni*ues and new antibiotics! "hese drugs, such as penicillin, are among the widel prescribed drugs against bacterial infections! Aith electrophoresis, e-perts are not onl able to s nthesize new antibiotics but are also able to anal ze which t pes of bacteria are antibiotic$resistant!

,. .accine !nalysis
Baccine anal sis is one of the man important applications of electrophoresis! "here are se'eral 'accines that ha'e been purified, processed and anal zed through electrophoresis, such as the influenza 'accine, hepatitis 'accine and polio 'accine! "he e-act steps done in the 'accine anal sis, howe'er, cannot be determined due to confidentialit reasons of the pharmaceutical companies! Ne'ertheless, data reports from 'accine manufacturers such as A eth, +erc% and &anofi$A'entis presents electrophoresis as an effecti'e 'accine anal sis method! References

4erg," mocz%o,&tr er!4iochemistr :9ifth Edition!New 7or%, New 7or%!A!,!9reeman and ;ompan ,1//1! http:CCwww!chemsoc!orgCE-emplar;hemCentriesC1//5CleedsDchromatog raph Cchromatograph C