Glycogen Determination from Tissues: Difference between revisions
Wrote initial protocol |
added glucose quantification protocol |
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* 50 mM Sodium Acetate, pH 4.8 | * 50 mM Sodium Acetate, pH 4.8 | ||
* Amyloglucosidease | * Amyloglucosidease | ||
* Glucose quantification kit (Wako Autokit Glucose Buffer Solution cat # 439-9091) | |||
* Glucose standard solution (500 mg/dL; Wako) | |||
==Protocol== | ==Protocol== | ||
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# Resuspend pellet in 200 uL of 50 mM Sodium Acetate, pH 4.8 | # 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 | # Add 0.3 mg/mL amyloglucosidase and place at 37C for 3h | ||
# Quantify glucose | # Quantify glucose using 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: | Reference: | ||
PMID 15282316 | PMID 15282316 | ||