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Glycogen Determination from Tissues

376 bytes added, 16:57, 6 May 2010
added glucose quantification protocol
* 50 mM Sodium Acetate, pH 4.8
* Amyloglucosidease
* Glucose quantification kit (Wako Autokit Glucose Buffer Solution cat # 439-9091)
* Glucose standard solution (500 mg/dL; Wako)
==Protocol==
# Resuspend pellet in 200 uL of 50 mM Sodium Acetate, pH 4.8
# Add 0.3 mg/mL amyloglucosidase and place at 37C for 3h
# Quantify glucoseusing kit:## Add 1-5 uL glucose standard for standard curve## Add 10 uL digested glycogen## Mix and incubate at 37C for 5 min## Measure absorbance at 505 nm## Calculate glycogen levels as umoles glucose released per g tissue (should be ~10)
Reference:
PMID 15282316