| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
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| Other Sizes |
| Targets |
Phenolphthalein beta-D-glucuronide targets beta-glucuronidase (GUS, EC 3.2.1.31), a lysosomal enzyme that cleaves beta-D-glucuronic acid moieties from various substrates. Upon enzymatic hydrolysis, the substrate releases phenolphthalein, which is colorless at acidic to neutral pH but becomes purple (λmax 550-555 nm) under alkaline conditions. The GUS gene from E. coli is commonly used as a reporter in plant and yeast transformation. The color change is specific for measuring GUS activity.
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|---|---|
| ln Vitro |
In vitro, Phenolphthalein beta-D-glucuronide (1-20 mM) is used as a substrate for beta-glucuronidase in a colorimetric assay. The GUS enzyme cleaves the glucuronide bond, releasing phenolphthalein. Upon addition of alkaline stop buffer (e.g., 2 M NaOH or 2 M glycine-NaOH, pH 10.4), the phenolphthalein anion develops a magenta color (A₅₅0). The reaction is linear with enzyme concentration and time. For purified E. coli GUS, the Km for the substrate is 0.2-0.5 mM; the specific activity is 1-10 umol phenolphthalein released/min/mg protein.
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| ln Vivo |
Not applicable; the compound is a substrate used in biochemical assays and is not used as a therapeutic in vivo. It can be used to detect GUS activity in plant tissues (ex vivo) or in bacterial cultures. It is not administered to animals for therapeutic purposes.
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| Enzyme Assay |
For GUS enzyme assays, E. coli GUS (0.1-1 U) is incubated with Phenolphthalein beta-D-glucuronide (0.2-10 mM) in 1× GUS assay buffer (50 mM NaH2PO4, pH 7.0, 10 mM beta-mercaptoethanol, 1 mM EDTA, 0.1% Triton X-100) at 37degC for 15-60 min. The reaction is stopped by adding stop buffer (2 M NaOH or 2 M glycine-NaOH, pH 10.4). Absorbance is measured at 550 nm. A standard curve of phenolphthalein (0.1-100 uM) is prepared. For kinetic analysis, the substrate concentration is varied (0.2-10 mM), and initial velocities are measured to determine Km and Vmax. For inhibitor studies (e.g., glucaro-1,4-lactone), the inhibitor is pre-incubated with the enzyme for 5 min before adding the substrate.
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| Cell Assay |
For cell-based GUS assays (plant cells or tissues), the sample is homogenized in GUS extraction buffer (50 mM NaH2PO4, pH 7.0, 10 mM beta-mercaptoethanol, 1 mM EDTA, 0.1% Triton X-100). The extract is centrifuged (12,000g, 10 min, 4degC). The supernatant is incubated with the substrate (2-5 mM final) at 37degC for 1-24 h, depending on GUS expression levels. The reaction is stopped with stop buffer, and A₅₅0 is measured. For histochemical GUS staining, plant tissues are incubated with 5-bromo-4-chloro-3-indolyl beta-D-glucuronide (X-Gluc) rather than phenolphthalein-based substrates. For mammalian cells, endogenous GUS activity can be measured in cell lysates using the same protocol.
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| Animal Protocol |
Not applicable; the compound is used for ex vivo biochemical assays, not in animals.
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| ADME/Pharmacokinetics |
The substrate is water-soluble (10 mg/mL in water) and is stable in solution at 4degC for weeks. For storage, the powder is stored at -20degC, protected from light. The released product (phenolphthalein) is a light- and pH-sensitive dye; stop buffer should be added immediately after the reaction. Not for IV use.
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| Toxicity/Toxicokinetics |
Low toxicity. Phenolphthalein is a known laxative (at high doses) and may be carcinogenic in rats (intestinal tumors). However, the amounts used in assays are tiny (umol). The glucuronide substrate itself is non-toxic. Use standard PPE (gloves, goggles, lab coat). Avoid skin contact. Not a drug.
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| References | |
| Additional Infomation |
Phenolphthalein glucuronide is the parent cpd
Phenolphthalein beta-D-glucuronide is a research-grade substrate for beta-glucuronidase (GUS) activity assays. It is not an FDA-approved drug. It is used in GUS reporter gene assays for plant molecular biology and in coliform detection kits (e.g., for water quality testing). For research use only. |
| Molecular Formula |
C26H22O10
|
|---|---|
| Molecular Weight |
494.45
|
| Exact Mass |
494.121
|
| CAS # |
15265-26-6
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| Related CAS # |
6820-54-8 (hydrochloride salt)
|
| PubChem CID |
3032634
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| Appearance |
Typically exists as solids at room temperature
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| Density |
1.584g/cm3
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| Boiling Point |
804.5ºC at 760mmHg
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| Flash Point |
276.5ºC
|
| Vapour Pressure |
2.53E-27mmHg at 25°C
|
| LogP |
1.126
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| Hydrogen Bond Donor Count |
5
|
| Hydrogen Bond Acceptor Count |
10
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| Rotatable Bond Count |
5
|
| Heavy Atom Count |
36
|
| Complexity |
805
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| Defined Atom Stereocenter Count |
5
|
| SMILES |
OC1C=CC(C2(OC(=O)C3=CC=CC=C23)C2C=CC(O[C@@H]3O[C@H](C(=O)O)[C@@H](O)[C@H](O)[C@H]3O)=CC=2)=CC=1
|
| InChi Key |
FXJYOZKDDSONLX-XADSOVDISA-N
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| InChi Code |
InChI=1S/C26H22O10/c27-15-9-5-13(6-10-15)26(18-4-2-1-3-17(18)24(33)36-26)14-7-11-16(12-8-14)34-25-21(30)19(28)20(29)22(35-25)23(31)32/h1-12,19-22,25,27-30H,(H,31,32)/t19-,20-,21+,22-,25+,26?/m0/s1
|
| Chemical Name |
(2S,3S,4S,5R,6S)-3,4,5-trihydroxy-6-[4-[1-(4-hydroxyphenyl)-3-oxo-2-benzofuran-1-yl]phenoxy]oxane-2-carboxylic acid
|
| Synonyms |
Phenolphthalein β-D-glucuronide
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.0224 mL | 10.1122 mL | 20.2245 mL | |
| 5 mM | 0.4045 mL | 2.0224 mL | 4.0449 mL | |
| 10 mM | 0.2022 mL | 1.0112 mL | 2.0224 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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