AMPPD (Lumi-Phos Plus) (Synonyms: Lumi-Phos Plus; Lumigen PPD; PPD) |
Katalog-Nr.GC30019
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AMPPD (Lumi-Phos Plus), ein 1,2-Dioxo-Cyclohexan-Derivat, ist ein biochemisches ultrasensitives alkalisches Phosphatase-Substrat.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 122341-56-4
Sample solution is provided at 25 µL, 10mM.
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AMPPD (Lumi-Phos Plus) is a 1,2-dioxetane derivative that serves as an ultrasensitive chemiluminescent substrate for alkaline phosphatase and is widely used in biochemical detection [1-2]. Under the catalysis of alkaline phosphatase, AMPPD undergoes dephosphorylation to form an unstable intermediate. This intermediate decomposes, leading to cleavage of the dioxetane structure and emission of light at a wavelength of 470nm, with the luminescence intensity being proportional to the concentration of alkaline phosphatase[3-4].
References:
[1] Yue L, Liu YJ. Mechanism of AMPPD Chemiluminescence in a Different Voice. J Chem Theory Comput. 2013 May 14;9(5):2300-12.
[2] Tu L, Wang Y, Yang Y, et al. Spectroscopic study of the authentic emitter of AMPPD chemiluminescence in alkaline aqueous solution. Phys Chem Chem Phys. 2010 Jul 7;12(25):6789-94.
[3] Cate RL, Ehrenfels CW, Wysk M, et al. Genomic Southern analysis with alkaline-phosphatase-conjugated oligonucleotide probes and the chemiluminescent substrate AMPPD. Genet Anal Tech Appl. 1991 May;8(3):102-6.
[3] Cate RL, Ehrenfels CW, Wysk M, et al. Genomic Southern analysis with alkaline-phosphatase-conjugated oligonucleotide probes and the chemiluminescent substrate AMPPD. Genet Anal Tech Appl. 1991 May;8(3):102-6.
[4] Hai Z, Li J, Wu J,, et al. Alkaline Phosphatase-Triggered Simultaneous Hydrogelation and Chemiluminescence. J Am Chem Soc. 2017 Jan 25;139(3):1041-1044.
Alkaline Phosphatase Activity Assay
This protocol is adapted from research data and provided for reference only. Adjustments may be necessary based on specific experimental requirements.
(1) Enzyme Sample Preparation: Preserve samples containing alkaline phosphatase (e.g., cell lysate supernatant or immunodetection complexes) in 50mM carbonate buffer (pH=9.5) at 4°C for later use.
(2) Alkaline Phosphatase Activity Detection: The reaction system contains 0.0125–0.4mM AMPPD, 50 mM carbonate buffer (pH=9.5), and 1mM MgCl₂, with the total volume set according to the experiment. Incubate at room temperature for 20 minutes before detection.
(3) Photometer Detection: Use a luminometer to measure the chemiluminescent signal, recording either the average or integrated light intensity over 10 or 30 seconds.
(4) Film Imaging Detection: For alkaline phosphatase immobilized on membranes (e.g., Western blot or dot blot), incubate the membrane with AMPPD substrate and then expose to photographic film for imaging.
Precautions:
1) The optimal pH for the chemiluminescent reaction is 9.0–9.5, where the signal-to-noise ratio (enzymatic reaction/non-enzymatic hydrolysis) is highest.
2) For your safety and health, please wear a lab coat and disposable gloves when operating.
References:
[1] Bronstein I, Edwards B, Voyta JC. 1,2-dioxetanes: novel chemiluminescent enzyme substrates. Applications to immunoassays. J Biolumin Chemilumin. 1989 Jul;4(1):99-111.
| Cas No. | 122341-56-4 | SDF | |
| Überlieferungen | Lumi-Phos Plus; Lumigen PPD; PPD | ||
| Canonical SMILES | COC1(C2=CC(OP(O)(O)=O)=CC=C2)C3(C(C4)CC5CC4CC3C5)OO1 | ||
| Formula | C18H23O7P | M.Wt | 382.34 |
| Löslichkeit | Water : ≥ 125 mg/mL (326.93 mM) | Storage | Store at -20°C |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 2.6155 mL | 13.0774 mL | 26.1547 mL |
| 5 mM | 523.1 μL | 2.6155 mL | 5.2309 mL |
| 10 mM | 261.5 μL | 1.3077 mL | 2.6155 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid 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 master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL saline, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >98.00% Appearance: A solid
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 24 reference(s) in Google Scholar.)