Anthranilic acid
It is a white crystalline solid with a distinctive odor and is an important intermediate in the synthesis of various pharmaceuticals, dyes, and other industrial chemicals.
Anthranilic acid plays a crucial role in the metabolism of tryptophan and is involved in the production of niacin, a essential B-vitamin.
It has been studied for its potential therapeutic applications in areas such as pain management, inflammation reduction, and neurological disorders.
Researchers can leverage PubCompare.ai to optimzie their anthranilic acid-related research, locating the best protocols from literature, preprints, and patents using AI-driven comparisons to enhance reproducibility and accuracy, and streamline their research workflow to achieve greater results.
Most cited protocols related to «Anthranilic acid»
Complete amplification of the viral genome was performed using a PCR reaction with Phusion High Fidelity (Thermo Fisher) enzyme and the designed primers ZIKV-Forward (5′ CG ATT AAG TTG GGT AAC GCC AGG GT 3′) and ZIKV-Reverse (5′ T AGA CCC ATG GAT TTC CCC ACA CC 3′). The PCR product containing SP6 promoter followed by complete viral cDNA was purified with the DNA clean and concentration kit (Zymo Research). In vitro transcription was performed using the RiboMAX™ Large-scale RNA Production Systems kit (Promega), as instructed by the manufacturers.
encoding human TRPM2 (8 (link)) with a
C-terminal EE epitope (29 ) was
used. Mutations were introduced using QuikChange system (Stratagene) and
confirmed by sequencing. The constructs encoding subunit concatemers with a
C-terminal EE epitope were made as follows. Firstly, the sequence (nucleotides
1–700) encoding part of the TRPM2 N terminus (TRPM2N) was amplified by
PCR using Pfu and forward primer
5′-TC
sequence underlined and TRPM2 sequence in italic) and reverse primer
5′-TCAGTACAGGTAGAGCAAGGTGTCC-3′. The resultant PCR product
containing XbaI site was inserted into pCR2.1 following the manufacturer's
instructions (Invitrogen) to generate TRPM2N-pCR2.1. Secondly, the vector
sequence between EcoRI and XbaI and the TRPM2 sequence between XbaI and SacI
were separately excised from TRPM2N-pCR2.1 and ligated with the sequence
between SacI and EcoRI from TRPM2-EE-pcDNA3.1 to generate TRPM2-EE-pCR2.1.
Finally, the TRPM2-EE sequence between XbaI and HindIII was excised from
TRPM2-EE-pCR2.1 to replace the sequence between XbaI and PmeI in
TRPM2-Myc-pcDNA3.1 to produce the constructs encoding concatenated subunits
(see
Maintenance of human embryonic kidney cells (HEK293) and transient
transfection with plasmids were described previously
(29 ).
Biotin Labeling and Western Blotting Analysis—Experiments
were performed as described previously
(29 ,
30 (link)). Proteins were resolved on
SDS-PAGE gels and detected using primary rabbit anti-EE antibody (1:2000
dilution; Bethyl Laboratories) and secondary goat horseradish
peroxidase-conjugated anti-rabbit IgG antibody (1:2000 dilution; Santa Cruz
Biotechnology).
Electrophysiological Recording—Whole-cell recordings were
made using an Axopatch 200B amplifier at room temperature 24–48 h after
transfection as described previously
(29 ,
30 (link)). The data were filtered at
2 kHz and sampled at 10 kHz. Cells were held at –40 mV and voltage ramps
with a 1-s duration from –120 mV to 80 mV were applied every 5 s. The
currents at –80 mV denoted in the figures by circles (see Figs.
were obtained from the current responses to voltage ramps. In some
experiments, cells were held constantly at –40 mV or 40 mV, and the
currents were plotted as continuous lines (see
Intracellular solution contained (in m
MgCl2, 10 HEPES, 1 Na2ATP, and 1 ADPR.
of extracellular solutions used. Flufenamic acid (FFA) (0.5 m
(31 (link)) or
N-(p-amylcinnamoyl) anthranilic acid (20 μ
(32 (link)) was applied at the end of
each recording via a RSC-160 system (Biologic Science Instruments) to confirm
TRPM2 channel-mediated currents.
m
Standard | 147 | 2 | 1 | 2 | 10 | 13 |
147 NaCl + 2 CaCl2 + 1 MgCl2 | 147 | 0 | 1 | 2 | 10 | 13 |
147 NaCl (no CaCl2) | 147 | 0 | 0 | 0 | 10 | 24 |
147 NaCl + 0.1 CaCl2 | 147 | 0 | 0 | 0.1 | 10 | 24 |
147 NaCl + 0.3 CaCl2 | 147 | 0 | 0 | 0.3 | 10 | 22 |
147 NaCl + 1 CaCl2 | 147 | 0 | 0 | 1 | 10 | 21 |
147 NaCl + 2 CaCl2 | 147 | 0 | 0 | 2 | 10 | 17 |
147 NaCl + 3 CaCl2 | 147 | 0 | 0 | 3 | 10 | 11 |
147 NaCl + 10 CaCl2 | 147 | 0 | 0 | 10 | 10 | 0 |
147 NaCl + 2 MgCl2 | 147 | 0 | 2 | 0 | 10 | 20 |
110 CaCl2 | 0 | 0 | 0 | 110 | 10 | 24 |
110 MgCl2 | 0 | 0 | 110 | 0 | 10 | 24 |
determined from current responses to the aforementioned voltage ramps. After
cell-attached configuration was established in standard extracellular
solution, application of voltage ramps started and continued throughout
experiments, during which whole-cell configuration was achieved at least 2 min
after the external solution was replaced with the indicated extracellular
solutions. The flufenamic acid/anthranilic acid-insensitive current components
were negligible (e.g.
1B
The reversal potentials were corrected for liquid junction potentials as we
described previously (33 (link)). Ion
activities were used, converted from ion concentrations using the following
coefficients: γNa = 0.75, γCa = 0.28, and
γMg = 0.34. The relative permeability
PX/PNa (X = calcium or
magnesium) were derived using the Goldman-HodgkinKatz equation
(8 (link),
23 (link)):
PX/PNa = [Na]i exp(ErF/RT)(1 + exp
(ErF/RT))/4[X]o,
where F, R, and T are Faraday constant, gas constant, and
absolute temperature.
Data Analysis—All the data, where appropriate, are presented
as mean ± S.E. The calcium inhibition was estimated by fitting to the
Hill equation: I/I (%) = (100–C)/[1+
([calcium]/IC50)n], where I is the
sustained current as a percentage of the peak current (Ip),
C is the Ca2+-insensitive current component,
IC50 is the concentration producing half-maximal inhibition, and
n is the Hill coefficient. Curve fitting was carried using Origin
(OriginLab, Northampton, MA). Comparisons were made using the Student's
t test between two groups or analysis of variance (post hoc Tukey)
between multiple groups with significance at the level of p <
0.05.
Concurrent analysis of tryptophan, kynurenine, 3-hydroxykynrenine (3-HK), 3-hydroxyanthranilic acid (3-HAA), and anthranilic acid (AA) was performed with UHPLC as described by Jones et al.16 (link), using an injection volume of 20 μL of the prepared extract from each samples. KA detection was performed using a gradient mobile phase comprise of 50 mM sodium acetate buffer supplement with 25 mM zinc acetate (dihydrate) to enhance fluorescence intensity and 2.25% acetonitrile as organic modifier (Solvent A), and 10% acetonitrile (Solvent B). Each sample (10 μL) was injected into a Poroshell RRHT C-18, 1.8 μm 2.1 × 100 mm column (Agilent Technologies, Inc, Santa Clara, CA) maintained at 38 °C for 12 min run time at a unison flowrate of 0.75 mL/min. The gradient elution consisted of 100% solvent A for 3 min and then 50% solvent A and 50% solvent B for 2 min, followed by 100% B for 2 min and 100% solvent A (run time 10 min). This gradient ensures sufficient time for KA retention while minimizing potential build-up of pressure due to precipitation of the high salt buffer. Detection of KA used fluorescence (excitation and emission wavelengths of 344 and 388 nm, respectively with a retention time of 1.5 min). Agilent OpenLAB CDS ChemStation (Edition C.01.04) was used to analyze the chromatograms (
For GCMS, 50 μL of the prepared extract were derivatized. Concurrent analysis of PA and QA were carried out as described by Smythe et al.17 (link) with slight modification using an Agilent 7890 A GC system coupled with Agilent 5975 C mass spectrometry detector and Agilent 7693 A autosampler (Agilent Technologies, Inc, Santa Clara, CA) with one microliter of derivatized mixture. Separation of PA and QA were achieved with a DB-5MS column, 0.25 μm film thickness, 0.25 mm × 30 m capillary column (Agilent Technologies, Inc, Santa Clara, CA) within 7 min but the assay run time was set for 12 min to prevent sample carryover. Concentrations of PA and QA were analyzed using Agilent GC/MSD ChemStation software (Edition 02.02.1431) and interpolated from the established six-point calibration curves based on the abundance count ratio of the metabolites to their corresponding deuterated internal standards within each standards and samples (
The intra- and inter-assay CV was within the acceptable range of 4–8% for UHPLC assays and 7–10% for GCMS assays calculated from the repeated measures of the metabolites standards incorporated during the sequence run. Nicotinamide adenine dinucleotide (NAD+) was measured with 20 μL of neat serum using a previously described method18 (link).
Most recents protocols related to «Anthranilic acid»
Proton nuclear magnetic resonance spectroscopy analysis ( 1 H-NMR) was conducted to confirm the labelling. For this purpose, fluorescently labelled mannose and unlabelled mannose were dissolved in deuterated water (D 2 O) obtained from Sigma Aldrich (St Louis, MO, USA) at 200 mM, while anthranilic acid was dissolved in deuterated dimethyl sulfoxide (DMSO-D6; Cambridge Isotope Laboratories, Inc. (Andover, MA, USA)) at the same concentration. The solutions were then placed in NMR tubes (600 µl) to be characterized using a Bruker NEO-750 equipment (Karlsruhe, Germany). (
Example 1
Procedure:
Approximately 90% of the total volume of Water for Injection (WFI) was added to a preparation vessel. The WFI was deoxygenated. Tetrofosmin, reducing agent selected from stannous chloride dihydrate, transchelator selected from Sodium D-gluconate, radioprotectant selected from Gentisic acid, para-amino benzoic acid, maleic acid, anthranilic acid and combinations thereof and sodium hydrogen carbonate were dissolved. The solution obtained was deoxygenated and final volume and the bulk solution was deoxygenated. The solution was sterile filtered. The vials were partially stoppered and then lyophilized.
Top products related to «Anthranilic acid»
More about "Anthranilic acid"
It is a white crystalline solid with a distinctive odor and is an important intermediate in the synthesis of various pharmaceuticals, dyes, and other industrial chemicals.
Anthranilic acid plays a crucial role in the metabolism of tryptophan, an essential amino acid, and is involved in the production of niacin, a vitamin B-complex.
Researchers have explored the potential therapeutic applications of anthranilic acid in areas such as pain management, inflammation reduction, and neurological disorders.
Closely related compounds like gibberellic acid, jasmonic acid, and thymine-d4 have also been studied for their biological and chemical properties.
Amino acids like glutamic acid-d5 and tryptophan-15N2 are often used in conjunction with anthranilic acid for various analytical and experimental purposes.
Leveraging the power of PubCompare.ai, researchers can optimize their anthranilic acid-related studies by locating the best protocols from literature, preprints, and patents using AI-driven comparisons.
This can enhance the reproducibility and accuracy of their experiments, while also streamlining their research workflow to achieve greater results.
Whether exploring the metabolism of anthranilic acid or developing new pharmaceutical applications, PubCompare.ai can be a valuable tool in the researcher's arsenal.