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alpha-D-Glucose-1-Phosphate (G1P)
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Glucose-1-phosphate can be used as a substrate in the assay of the activities of phosphoglucomutase and UDP-glucose pyrophosphorylase and also in the preparation of other sugar phosphates.
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1.Source: |
Disodium, tetrahydrate
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Form: |
Crystalline powder
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Molecular Weight: |
376.1
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Solubility: |
Distilled water or dilute buffer
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Stability: |
Store at 4° C (39° F)
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Protein: |
99-100% glucose-1-phosphate (assayed enzymatically)
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Catalog No.: |
101J0000
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Glucose-1-P is converted to Glucose-6-P with phosphoglucomutase (EC 2.7.5.1). Glucose-6-P is oxidized by
NADP+ and glucose-6-P dehydrogenase (EC 1.1.1.49) to 6-phosphogluconate (Methods of Enzymatic Analysis,
Bergmeyer, H.U. ed. VOL 3, 1233, 1974, Academic Press, New York).
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- 0.1 M Triethanolamine buffer containing 2 mM EDTA and 5 mM MgCl2, pH 7.6.
- 0.02 M NADP+ (17.4 mg/ml) in distilled water.
- Phosphoglucomutase (PGM). Prepare in buffer to yield a final concentration of 100 U/ml. Prepare fresh prior to assay.
- Glucose-6-P-dehydrogenase (G6PDH). Prepare in buffer to yield a final concentration of 100 U/ml. Prepare fresh prior
to assay.
- 0.012 M Glucose-1-P (substrate). Dissolve 44.6 mg glucose-1-P, dipotassium dihydrate in 10 ml distilled water. Prepare
fresh.
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- Set spectrophotometer (equipped with a strip chart recorder and temperature control) at 37°C and 340 nm.
- Into a cuvette, pipette the following in the amounts indicated:
Triethanolamine buffer with EDTA and MgCl2 2.70 ml NADP+ 0.10 ml NADH
0.10 ml Glucose-6-P-DH 0.10 ml Glucose-1-P (substrate) 0.01 ml - Incubate cuvette in the spectrophotometer at 37°C for 5 minutes.
- Record the initial absorbance at 340 nm (Ai).
- Initiate the reaction by adding 0.10 ml of phosphoglucomutase solution to the cuvette.
- Follow the reaction for 10 minutes. Record the final absorbance at 340 nm (Af).
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