Glycogen Assay (via Gustavo)
1.
Cut and weight tissues
2.
Add to 1.0 ml of 30% KOH w/Na2SO4. Make sure that the tissue is
completely immersed in the solution.
3.
Boil tubes with foil (to avoid evaporation) for 20-30’ until complete digestion
(sharke tubes frequently)
4.
Vortex and cool in ice.
5.
Add 2.0 ml of 95% EtOH to preciptate glycogen from alkaline digestate.
6.
Vortex and incubate samples in ice for 30 min
7.
Centrifuge tubes at 550 g for 30 min
8.
Carefully pour supernatant and dry tubes upside down (5 min)
9.
Re-dissolve pellet (glycogen) in 1 ml of ddH20 (vortex until goes into
solution).
10.
Have the glycogen standards ready (0, 25, 50, 75 and 100 g/ml)
11.
Add 1 ml of 5% Phenol

Make sample duplicates: muscle 400l (300 l) + 600 l (700 l) ddH20,
Liver 50l (25l) + 950l (975l) ddH2O










Rapidly repipette 5ml of 96-98% H2SO4 direct stream to the tube content.
Incubate on ice bath for 30min
Have cuvettes ready for OD
Prepare blank with 1ml of ddH2O
Read OD at 490 nm (within 30min)