Polyacrylamide Gel Electrophoresis- PAGE - Amrita University

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PAGE (Polyacrylamide Gel Electrophoresis), is the most widely used analytical method to resolve separate components of a protein mixture based on size.
For this protein molecules of different shapes and sizes, need to be denatured. This is done with the aid of SDS, so that the proteins no longer have any secondary, tertiary or quaternary structure. The proteins covered by SDS are negatively charged. When loaded onto a gel matrix and placed in an electric field, the proteins will migrate towards the anode (positively charged electrode). They are then separated by a molecular sieving effect based on size. After visualization by a protein-specific staining technique, the size of a protein can be estimated by comparison of its migration distance with that of a known molecular weight marker.

[@TheChC47]

@ktochi297 i'm not 100% sure, but i think that the advantage of Tris is that it's isotonic and its pH is close to the physiological pH (7). So, it makes sense a Tris pH 6.8 for the stacking instead of 8.8. But i'm just wondering :P

[@safeyyasofy5515]

very good explained !! well done :)

[@fitriamfauzi2586]

how is the connection between sds chamber and power suply is not correct? what is the effect with the sample or the gel?

[@Drshinemadathil]

thank u.......

[@kbnsahebmaski8857]

tnx its helpful to meeee

[@majeedaraza81]

stop using music behind

[@langitlama8015]

thank you :)

[@LFS2333]

very nice

[@akand999]

For Stacking gel, its Tris of pH 6.8, urs is wrong... check it again

[@ktochi297]

@akand999 I think the pH of the stacking and resolving gels would depend on what your protein sample is.

[@MsRicha01]

I AM PLANNING TO PURCHASE THIS SYSTEM. CAN PEOPLE WHO USED THIS LET ME KNOW IF THIS IS GOOD OR NOT. WE ARE CURRENTLY USING HOEFFER SE260. GOOD SYSTEM BUT GEL CASTING IS BIT DIFFICULT. I HAD HEARD SOMEWHERE THAT BIORAD GEL CASTING LEAKS. PLEASE GUIDE

[@Anupriya014]

nice presentation.. :) thnx

[@RyanRamanayake]

The accent needs to be a bit more clear, all info is right and presented very well :)

[@NetrapalSinghShekhawat]

Nice

[@somyaupadhyay8388]

nice dna bands are formed