Glycogen Determination from Tissues: Difference between revisions
m Added step 2,clarified how to mix sample, added clarification to step 3 as to ensure all sample is immersed in KOH, added step 6, clarified the washing steps in order, clarified how to dry pellet |
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# Resuspend pellets in 200uL amyloglucosidase solution and incubate at 37C for 3h-O/N | # Resuspend pellets in 200uL amyloglucosidase solution and incubate at 37C for 3h-O/N | ||
# Quantify glucose using kit: | # Quantify glucose using kit: | ||
## Add | ## Add 100 uL of Glucose buffer solution to each well, including wells for standard curve and blank | ||
## Add | ## Add 1-5 ul of glucose standard 200mg/dL diluted 1:5 | ||
## Add 10 uL digested glycogen | ## Add 10 uL digested glycogen for fasted samples, and use a 10X dilution for refed tissues (using either 5 or 2ul from the diluted sample) | ||
## Mix and incubate at 37C for 5 min | ## Mix and incubate at 37C for 5 min | ||
## Measure absorbance at 505 nm | ## Measure absorbance at 505 nm | ||