Hi,I currently work for a Biopharmaceutical company and I would like to find out about the SDS PAGE validation.(How to go about validating the test,what should be taken into consideration,methodology,gel, etc)
Regards
ThatoK
You should always follow the standard concentrations that is given as per your references for making the Loading gel and Resolution gel. Different products have different concentrations.
Validation must start right here
[[COLOR=“blue”]B]- First write a SOP what you want to perform.
-Compare the Gels with different concentrations with minute changes
-Use standard molecular weight markers and evaluate the results
-Compare with International Pure standards
-Compare with your pure products
-Compare your products based on different quantities you load on your gel.
-Always keep the elctricity constant for all runs
-Prepare the Gel buffers and make a standard SOP’s stating the quality of chemicals and water to make such buffers.
-Make sure that you run the gel always with a Marker or Comperator.[/b][/color]
Once this procedure is done and you fix the strengths of gels , Buffers and quantities of samples you load --You must run 3 batches of Gels under standard conditions you define, Take the Gels and compare each one on a Gel Documentor.
Your error should be less than 1% between the runs.
Some errors happen when inexperienced hands load the gels with micropippete.This causes either loading more, Pumping air bubbles or disturbing the gel loading cavities/Slots.
A trained hand is necessary or you need an expert supervision.
Ensure that you do not have leaks and take care that you do not have any airbubbles in the interphase of Loading Gel and Resolution gel.
Most folks use IsoButyl alcohol for such needs so that entrapment of air is prevented.
Regards