Electrophoresis refers to the migration of charged solutes or particles of any size in a liquid medium under influence of an electrical field. Depending on the charge on the molecules, they move towards either anode or cathode.

Electrophoresis of positively charged particles (cations) is sometimes called cataphoresis, while electrophoresis of negatively charged particles (anions) is sometimes called anaphoresis.

Required reagent for protein electrophoresis

Hippurate buffer pH 8.8

1% Agarose gel

BPB(bromophenol blue) dye

Amido black stain

Methanol

5% glacial acetic acid

Sample Preparation

Steps of Serum protein electrophoresis

Gel preparation

1. Clean the glass slide with tape water and and dry it with tissue paper.
2. Cut a rectangle four borders from X-ray plate and Stick X-ray plate with feviquick to the periphery of glass slide so the thickness of gel is same as thickness of X-ray plate.
3. Prepare 1% Agarose and pour hot agarose gel to non siliconized rough glass slide.
4. Cover the gel with siliconized glass slide carefully without entrapping air bubbles.
5. Allow the gel to get solidify for 10 minutes.
6. After 10 minutes removes siliconized glass slide by sliding movement and take care so that gel does not broken down.
7. Gel is ready for sample application.
8. Immediately apply the sample to prevent excess drying.

Sample Applicator Preparation Prepare comb from the stainless steel as shown in image. Attach the magnetic strip to the ruler. Stick the ruler with the platform by feviquick. Stick the Applicator to magnetic strip. Applicator can be move up and down.

Sample application

1. Put Rough non sliconized glass slide below the sample applicator.
2. Apply 4 micro liter sample over the sample applicator at the edge of applicator as shown in figure. so that excess sample will be removed on glass slide.
3. Lift up the sample applicator and wipe the back side of applicator so that backward running of sample can be prevented.
4. Replace the rough glass slide with glass slide having prepared gel.
5. Apply the sample near the one end of gel with the applicator and wait for 2 minutes to allow the samples to get completely absorbed within the gel.
6. Remove the applicator.