| Size | Price | Stock | Qty |
|---|---|---|---|
| 100mg |
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| 250mg |
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| 500mg | |||
| Other Sizes |
| Targets |
Acid Phosphatase [1]
Alkaline Phosphatase [2] |
|---|---|
| ln Vitro |
Advice (This is our suggested protocol, which should be adjusted to suit your particular needs as it only offers guidance). Assay for serum acid phosphatase[1]: 1. Mix 50 μL of 4-methylumbelliferate phosphate (5.0 mM), 10 μL of 1.0 M pH 6.0 buffer, and 5.0 μL of serum enzyme into a solution to make 100 μL. 2. Include the following reagents (final concentrations): 45 mM EDTA, 6 mM EGTA, 4 mM 1,10-phenanthroline, 6 mM 8 quinolinol, 6 mM 8-hydroxy-5-quinolinesulfonic acid, 20 mM sodium tartrate, 15 mM sodium fluoride, and 3000 mM 2-mercaptoethanol. 3. To stop the reaction, add 2.9 mL of 0.1 M ammonium hydroxide/glycine buffer (pH 10.5) to the reaction mixture and incubate it for 15 minutes at 37°C. 4. Using a fluorometer, measure the fluorescence after thoroughly mixing the sample.
- 4-Methylumbelliferyl phosphate is a fluorescent substrate for acid phosphatase. It was used to determine serum acid phosphatase activity in patients with Gaucher's disease. The hydrolysis of the compound by acid phosphatase releases 4-methylumbelliferone, which exhibits fluorescence (excitation: ~360 nm, emission: ~440 nm). The assay showed good linearity over the enzyme activity range of 0.1-10 U/L, with a detection limit of 0.05 U/L. Serum acid phosphatase levels in Gaucher's disease patients (mean: 8.3 U/L) were significantly higher than those in healthy controls (mean: 2.1 U/L) [1] - 4-Methylumbelliferyl phosphate serves as a substrate for alkaline phosphatase isoenzymes. When used in cellulose acetate membrane electrophoresis, it enabled the separation and quantification of alkaline phosphatase isoenzymes (e.g., liver-type, bone-type) from serum. The hydrolysis product's fluorescence allowed direct visualization and densitometric analysis of isoenzyme bands, with a linear detection range of 0.2-15 U/L for total alkaline phosphatase [2] |
| Enzyme Assay |
- Acid phosphatase activity assay (serum): Serum samples were diluted in acetate buffer (pH 5.0). 4-Methylumbelliferyl phosphate was added to a final concentration of 0.5 mM, and the mixture was incubated at 37°C for 30 minutes. The reaction was terminated by adding glycine-NaOH buffer (pH 10.3). Fluorescence intensity was measured at excitation 365 nm and emission 450 nm, and acid phosphatase activity was calculated using a 4-methylumbelliferone standard curve [1]
- Alkaline phosphatase isoenzyme assay: Serum samples were electrophoresed on cellulose acetate membranes using barbital buffer (pH 8.6). The membrane was incubated with a solution containing 4-Methylumbelliferyl phosphate (1 mM) and Tris-HCl buffer (pH 9.8) at 37°C for 15 minutes. After washing, the membrane was exposed to UV light (365 nm) to visualize fluorescent bands corresponding to alkaline phosphatase isoenzymes. Densitometric scanning was used to quantify the relative activity of each isoenzyme [2] |
| References |
|
| Additional Infomation |
4-Methylumbelliferone phosphate is a synthetic fluorescent substrate widely used for the quantitative detection of phosphatase activity[1][2]. Its core mechanism is that phosphatases (acidic or alkaline) hydrolyze the substrate to release the fluorescent product 4-methylumbelliferone, thereby enabling sensitive and specific quantification of enzyme activity by fluorescence[1][2]. It is a research and clinical diagnostic tool, not a therapeutic drug. Its main applications include the detection of serum acid phosphatase in the diagnosis of Gaucher disease[1] and the analysis of alkaline phosphatase isoenzymes for the differential diagnosis of liver/bone diseases[2]. This compound has high substrate specificity for phosphatases and minimal cross-reactivity with other hydrolases within the physiological pH range[1][2].
|
| Molecular Formula |
C10H9O6P
|
|---|---|
| Molecular Weight |
256.14866
|
| Exact Mass |
240.018
|
| CAS # |
3368-04-5
|
| Related CAS # |
4-Methylumbelliferyl phosphate disodium;22919-26-2
|
| PubChem CID |
65118
|
| Appearance |
White to off-white solid powder
|
| Density |
1.6±0.1 g/cm3
|
| Boiling Point |
513.6±60.0 °C at 760 mmHg
|
| Melting Point |
215-218 °C
|
| Flash Point |
264.4±32.9 °C
|
| Vapour Pressure |
0.0±1.4 mmHg at 25°C
|
| Index of Refraction |
1.628
|
| LogP |
0.75
|
| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
6
|
| Rotatable Bond Count |
2
|
| Heavy Atom Count |
17
|
| Complexity |
398
|
| Defined Atom Stereocenter Count |
0
|
| InChi Key |
BCHIXGBGRHLSBE-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C10H9O6P/c1-6-4-10(11)15-9-5-7(2-3-8(6)9)16-17(12,13)14/h2-5H,1H3,(H2,12,13,14)
|
| Chemical Name |
(4-methyl-2-oxochromen-7-yl) dihydrogen phosphate
|
| HS Tariff Code |
2934.99.9001
|
| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
H2O : ~20.83 mg/mL (~81.32 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 10 mg/mL (39.04 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
(Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.9040 mL | 19.5198 mL | 39.0396 mL | |
| 5 mM | 0.7808 mL | 3.9040 mL | 7.8079 mL | |
| 10 mM | 0.3904 mL | 1.9520 mL | 3.9040 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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