Sodium Doedecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS PAGE)

Hello everyone , 

Dixy is back with new experience to be shared  :)

Today I will be writing few important points about SDS PAGE Electrophoresis,  as I have used this technique many time during my Post graduation  . 

When I first performed SDS , that time I didn't knew  even to seal the apparatus , but now i can interdependently use it.  So don't be afraid of this technique , you can also do it easily by just following few points which I will be mentioning below.

 You must have seen the apparatus

 It consists of 2 plates, a comb,2 clamps, 2 side spacer, a buffer tank , power pack

Do you know you can prepare gel of different porosity by mixing  ingredients in different concentration 

• But the first thing is to do is sealing the apparatus with 1 % Agarose.
• All the step should be carried out with Gloved hands.
• All ingredient should be prepared fresh.
• Also point should be noted , to mix the ingredients gently , ensuring no air bubbles forms.
• Carefully pour the mixture of gel directly into glass plate assembly as soon the TEMED is added ,  not forgetting the fact that the mixture contains acrylamide and TEMED which are carcinogenic .
• Add water above the Gel mixture which allows polymerization of gel
• Once  comb is placed , don't move it , this way u will not get uniform wells.
• Fill the buffer in buffer tank 
• Always add sample in small volume with micropipette
• Cover the lid and connect the electrophoresis tank to the power supply.
• Allow the sample to run till it reaches 3/4 of gel
• while separating  gel  from plate , care should be taken not to tear the gel, or else u may lose separated  proteins.
• The gel is stained with Coomassie blue stain or Silver stain ,and destained 

Hope this may help you,

Thank you

With love ,

-Dixy