Context to curriculum whatsapp group discussions related to SDS -PAGE
Photo :Akhil Gupta
[17/04, 4:22 pm] Akhil : ID PAGE
[17/04, 4:24 pm] Akhil : ID SDS PAGE
One dimensional Sodium Dodecyl Sulfate polyacrylamide gel electrophoresis
[17/04, 4:24 pm] Akhil: One dimensional refers to protein separation based on molecular weight
[17/04, 4:32 pm] Arunan sir : Please give essential descriptions for each photo updates to inspire readers, who are from different backgrounds, but surely prepared to get excited. @~Akhil Gupta Samuel
[17/04, 4:33 pm] Arunan sir : What is PAGE, in simple terms?
We have school students onwards in the group. @~Akhil Gupta Samuel
Let’s inspire all.
[17/04, 4:34 pm] Arunan sir : Any examples of popular proteins and their molecular weight? @~Akhil Gupta Samuel @ayanasudheer @Theertha
[17/04, 4:42 pm] Akhil: PAGE is the technique used to seperate a mixture of proteins based on their molecular weight
Each protein is a garland of many amino acids✨ A. A✨( monomer of any protein )
Each protein has many amino acids (A. A )each with different charge
Some A. A are positively charged
Some are negatively charged
[17/04, 4:43 pm] Akhil: In SDS PAGE
SDS is a detergent which binds to the proteins in the mixture are denatures them
And gives all proteins a uniform negative charge
[17/04, 4:45 pm] Akhil: PAGE apparatus as shown in this picture has electrodes
Since through SDS we have given them negative charge
All proteins when applied electric feild in the apparatus will migrate towards
Postive electrode
[17/04, 4:52 pm] Akhil: The proteins are first loaded in the wells of the pre-made acrylamide gel
Acrymalide Gel
[17/04, 4:53 pm] Akhil: Acrylamide gel acts as a seive in seperating proteins
Smaller size proteins migrate faster down
Larger size dont
[17/04, 4:54 pm] Akhil: Its NOT LIKE CHROMATOGRAPHY seperation❌
Its seperation based on molecular weight in an applied electric feild
[17/04, 4:57 pm] akhil: Proteins are denatured by SDS which converts all the secondary and tertiary structures ( more folded structures) to primary structure ( protein as open unfolded form )
SDS attains uniform negative charge to all proteins so that when electric feild is applied they migrate to positive charged electrode
Proteins are denatured by SDS which converts all the secondary and tertiary structures ( more folded structures) to primary structure ( protein as open unfolded form )
SDS attains uniform negative charge to all proteins so that when electric feild is applied they migrate to positive charged electrode
Photo:Akhil
Photo Akhil
Gels can be pre-made and stored a day before loading the samples and running
Gel can be stained by coomaise brilliant blue stain to visualise the bands
Description by Akhil Gupta
