Category: Experimental Procedures
Author: Admin eLabJournal
Labels: Western blotting
|- Number of gels:
|- Percentage of acrylamide:
|- Volume of separation gel:
Setup the SDS-PAGE gel casting system and check for leakage with water.
Only add directly before casting the gel !!
Immediately cast the gel (leave 2-3 cm for the stacking gel) and
add 1 ml of butanol (water-saturated)
Allow the gel to polymerize for at least 1h
Remove the butanol, rinse with some dH20 and remove all liquid with some filter paper
Only add directly before casting the gel!!
Immediately pipet the stacking gel on top of the polymerized separating gel, insert a comb with the appropriate number of wells and allow the gel to polymerize for 1h
Load the samples on gel and run the gel at 130 V until the blue
loading dye reaches the bottom of the gel.
Acrylamide is highly neurotoxic!! Prevent any skin contact and always wear gloves!
Based on: Laemmli UK (1970). "Cleavage of structural proteins during the assembly of the head of bacteriophage T4". Nature 227 (5259): 680-685