N-Phenylnaphthalen-1-amine |
Catalog No.GD19545
One-Click Copy Product Info
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N-Phenyl-1-naphthylamine is a dye that fluoresces strongly when bound to the inner phospholipid bilayer of Gram-negative bacteria.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 90-30-2
Sample solution is provided at 25 µL, 10mM.
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N-Phenyl-1-naphthylamine is a dye that fluoresces strongly when bound to the inner phospholipid bilayer of Gram-negative bacteria. N-Phenyl-1-naphthylamine can be used to measure outer membrane permeability. N-Phenylnaphthalen-1-amine is a fluorescence probe for odorant-binding proteins (OBP) with a dissociation constant of 1.67 μM. N-Phenylnaphthalen-1-amine exhibits an excitation wavelength of 337 nM and an emission wavelength of 407 nM[1][2].
References:
[1]. Ban L, et al., Biochemical characterization and bacterial expression of an odorant-binding protein from Locusta migratoria. Cell Mol Life Sci. 2003 Feb;60(2):390-400.
[2]. Vanderlinde EM, et al. Mutation of the sensor kinase chvG in Rhizobium leguminosarum negatively impacts cellular metabolism, outer membrane stability, and symbiosis. J Bacteriol. 2012 Feb;194(4):768-77.
Guide (The following is our recommended protocol. This protocol is only a guide and should be modified according to your specific needs).
Odorant Binding Protein (OBP) Binding Affinity Assay[1]
1. Materials Preparation
Fluorescent probe: N-phenyl-1-naphthylamine, dissolved in methanol (MeOH) to prepare a 1 mM stock solution
Protein solution: 2 μM odorant binding protein (OBP), dissolved in 50 mM Tris-HCl buffer (pH 7.4)
Experimental temperature: 25 °C
Fluorescence spectrophotometer: equipped with a 1 cm quartz cuvette, excitation and emission slit widths set to 5 nm
Detection wavelengths: Excitation wavelength (λex): 337 nm; Emission wavelength (λem): 407 nm
2. Fluorescence Binding Assay Procedure
2.1 Place 2 μM odorant binding protein solution into the cuvette.
2.2 Gradually add the N-phenyl-1-naphthylamine stock solution (1 mM in methanol), mixing gently after each addition.
2.3 Set the final concentration range to 1-20 μM.
2.4 After each addition, immediately record the fluorescence emission spectrum (range 380-460 nm).
2.5 Measure and record the emission intensity at 407 nm for plotting the binding curve.
3. Data Analysis
3.1 Plot fluorescence intensity against the concentration of free N-phenyl-1-naphthylamine.
3.2 Use a Scatchard plot for linearization and calculation of the binding constant.
3.3 Assume a 1:1 binding ratio and 100% protein activity.
2.4 The dissociation constant (Kd) of N-phenyl-1-naphthylamine with loc-OBP1 is 1.67 μM.
4. Competitive Binding Assay
4.1 Use a fixed concentration of 2 μM odorant binding protein and 1 μM N-phenyl-1-naphthylamine.
4.2 Add test ligands (dissolved in methanol) at final concentrations typically ranging from 1-20 μM.
4.3 Monitor the decrease in fluorescence intensity and plot the competitive binding curve.
4.4 Calculate the IC50, and then convert it to the dissociation constant (Kd) of the test ligand.
| Cas No. | 90-30-2 | SDF | |
| Chemical Name | 1-Naphthalenamine, N-phenyl | ||
| Formula | C16H13N | M.Wt | 219.29 |
| Solubility | DMSO : ≥ 100 mg/mL (456.02 mM) | Storage | 4°C, protect from light |
| 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 | 4.5602 mL | 22.8009 mL | 45.6017 mL |
| 5 mM | 912 μL | 4.5602 mL | 9.1203 mL |
| 10 mM | 456 μL | 2.2801 mL | 4.5602 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: >99.50% Appearance: A solid
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 30 reference(s) in Google Scholar.)