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Handling and safety instruction for SDS-PAGE according to the
standard Laemmli gel method
Issued by: Madeleine Durbeej-Hjalt
Date: 2013-04-28
Applies for group: Muscle Biology, EMV
NB! ONLY purchase and use pre-made acrylamide gels and solutions - this removes the significant
hazard of inhaling the toxic powder form. Avoid ALL contact with acrylamide solutions or
polyacrylamide gels.
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Sample preparation is not included in this handling and safety instruction.
For adjustment of pH see separate handling and safety instruction: pH-justering i
vattenlösning.
If ready made gels are used, skip “Prepare and cast stacking and separating gels”.
1. Prepare buffers; separating gel buffer, stacking gel buffer, electrophoresis running buffer
and 5 x loading sample buffer
Weigh and dissolve SDS, TEMED and ammonium persulfate in fume hood or on a ventilated bench
with Plexiglas hood. Use lab coat, safety glasses and double, disposable nitrile gloves. Change gloves
frequently, especially if you suspect contamination. Remove gloves carefully without touching skin.
In case of power or ventilation failure or if an evacuation is necessary, close bottles and close the
front of the fume hood. In case of a long power or ventilation failure the lab must be evacuated.
For adjustment of pH see separate handling and safety instruction: pH-justering i vattenlösning.
2. Prepare and cast stacking and separating gels (2 full size 10% gels)
Use lab coat, safety glasses and double, disposable nitrile gloves. Change gloves frequently, especially
if you suspect contamination. Remove gloves carefully without touching skin. Work in fume hood.
Limit acrylamide work to designated areas. Coat work bench in fume hood with absorbent bench
paper prior to working with acrylamide to prevent contamination of non-disposable surfaces. Change
this paper layer frequently.
Use barrier/filter tips or disposable plastic transfer pipettes when aliquoting acrylamide solutions to
help prevent contamination of pipettes.
In case of power or ventilation failure or if an evacuation is necessary, close bottles and close the
front of the fume hood. In case of a long power or ventilation failure the lab must be evacuated.
3. Prepare the sample and load the gel
All handling of β-Mercaptoethanol must be done in fume hood or on ventilated bench with Plexiglas
hood. Use lab coat, safety glasses and double, disposable nitrile gloves. Keep containers tightly
closed.
H2O-saturated isobutyl alcohol is prepared in fume hood.
Loading samples via needle syringe. Potential for needle stick injury. Handle with care, do not resheathe needles.
In case of power or ventilation failure or if an evacuation is necessary, close bottles and close the
front of the fume hood. In case of a long power or ventilation failure the lab must be evacuated.
4. Run the gel (electrophoresis)
Ensure all equipment is in good working order, and that lid is always on electrophoresis tank when in
operation.
It is important to note that the voltage and currents used during electrophoresis are dangerous
and potentially lethal. Thus, safety should be an overriding concern. An unusually high or low
voltage for a given current (milliamper) might indicate an improperly made buffer or an electrical
leak in the chamber.
Inspect electrodes and cables for defects prior to use. Don’t use any electrophoresis equipment that
you suspect has an electrical fault or is damaged. Ensure electrophoresis chamber has a secure lid
which prevents accidental contact with the electrified buffer solution
Ensure area around electrophoresis chamber is free of spills or leaks prior and during the running of
your gel.
5. Clean and take care of waste
Use the same personal protective equipment as during the task.
All chemicals hazardous to health and environment, in this case especially ammonium persulfate,
acrylamide, β-Mercaptoethanol, TEMED and isobutyl alcohol, must be collected and be disposed of
as chemical waste are disposed, follow instructions here.
All disposable glass is disposed of as Sharp/Pointed. Wipe bench and fume cupboard with water.
Before sending glass ware to be washed it must be properly rinsed.
Wash hands thoroughly with soap and water.
6. In case of an accident
Make sure you know the location of spill kits, eyewashes, safety showers before starting the work!
Accidental leakage or spillage of ammonium persulfate, acrylamide, β-Mercaptoethanol, TEMED
and isobutyl alcohol, or mixtures
Evacuate contaminated area and signpost. Contact BMC Service for spillage kit (it contains
absorption material, breathing protection and protective gloves. Study instruction in kit.
NB! TEMED and isobutyl alcohol are highly flammable. Turn off any electrical device in case of
spillage.
Environmental precautions: Prevent further leakage or spillage if safe to do so. Do not let product
enter drains.
First aid measures
General advice: Consult a physician. Show this safety data sheet to the doctor in attendance.
If inhaled: Move person into fresh air. If not breathing, give artificial respiration. Consult a physician.
In case of skin contact: Wash off with soap and plenty of water. Consult a physician if needed.
In case of eye contact: Rinse thoroughly with plenty of lukewarm water for at least 15 minutes and
consult a physician immediately. Bring portable eye wash flask on the way to hospital.
If swallowed: Rinse mouth with water. Consult a physician immediately.
NB! Any incidents resulting in injury or large spills must be reported. Near misses (dangerous
situations not leading to an incident) should also be reported.