> Chemicals & Drugs > Organic Chemical > Isoamyl acetate

Isoamyl acetate

Isoamyl acetate is a colorless, flammable liquid with a characteristic banana-like odor.
It is commonly used as a flavoring agent in food and beverages, as well as a solvent in various industrial applications.
Isoamyl acetate is an ester formed from isoamyl alcohol and acetic acid.
It has a molecular formula of C7H14O2 and a moleular weight of 130.18 g/mol.
Isoamyl acetate can be found naturally in some fruits and fermented products, and is also synthetically produced.
It has a wide range of uses in the food, fragrance, and chemical industries.
Researchers can leverage PubCompare.ai's AI-driven platform to optimize their isoamyl acetate research, identifiying the mst reproducible and accurate protocols from literature, preprints, and patents.

Most cited protocols related to «Isoamyl acetate»

Comparing DNA Extraction Methods for Human Microbiome

Cited 53 times

Six DNA extraction methods (Table 1) were compared in this study, representing different kinds and combinations of cell lysis mechanisms and DNA purification methods commonly used in the published literature on the human microbiome. Each method was evaluated using all 11 type strains and a mock community sample. The isolated genomic DNA was in a final volume of 200 µl.
Method 1. The QIAamp DNA mini kit (Qiagen, Valencia, CA) was used in this method with minor modifications. Briefly, 6 µl mutanolysin (25 KU/ml, Sigma-Aldrich) was added to a 500 µl aliquot of cells and the mixture was incubated for 30 min at 37°C. After this, 50 µl Proteinase K (20 mg/ml) and 500 µl AL buffer (Qiagen, Valencia, CA) were added and the sample was incubated for 30 min at 56°C. Then, 500 µl of ethanol was added and DNA was purified by using the columns provided in the kit (Qiagen, Valencia, CA) according to the manufacturer's instructions.
Method 2. A two-step cell lysis procedure was employed before use of the QIAamp DNA mini kit (Qiagen, Valencia, CA). First, 50 µl lysozyme (10 mg/ml, Sigma-Aldrich), 6 µl mutanolysin (25 KU/ml, Sigma-Aldrich), and 3 µl lysostaphin (4000 U/ml, Sigma-Aldrich) were added to a 500 µl aliquot of cell suspension followed by incubation for 1 hour at 37°C. Second, 600 mg of 0.1-mm-diameter zirconia/silica beads (BioSpec, Bartlesville, OK) were added to the lysate and the microbial cells were mechanically disrupted using Mini-BeadBeater-96 (BioSpec, Bartlesville, OK) at 2100 rpm for 1 minute. Further isolation and purification of the total genomic DNA from lysates was done using QIAamp DNA mini kits (Qiagen, Valencia, CA).
Method 3. Genomic DNA was extracted by using the QIAamp DNA stool kit (Qiagen, Valencia, CA) with a 95°C lysis step according to the manufacturer's instructions. Briefly, 500 µl ASL buffer was add to a 500 µl aliquot of cells suspension and the mixture was heated for 5 min at 95°C. Then, 100 µl Proteinase K (20 mg/ml) and 1 ml AL buffer were added and the mixture was incubated for 10 min at 70°C. After this, 1 ml of ethanol was added and the rest of the isolation protocol was continued as described by the manufacturer.
Method 4. A 210 µl aliquot of 20% SDS, 500 µl of a mixture of phenol∶ chloroform∶ isoamyl alcohol (25∶24∶1)], and 600 mg of 0.1-mm-diameter zirconia/silica beads (BioSpec, Bartlesville, OK) were add to a 500 µl aliquot of cells suspension. Microbial cells were then disrupted by using Mini-BeadBeater-96 (BioSpec, Bartlesville, OK) set on 2100 rpm for 1 min. Next, the mixture was centrifuged at full speed (14000 rpm) for 5 min to separate phases. The top aqueous layer was transferred to a clean 2 ml micro-centrifuge tube. Then, 0.1 volume of 3 M sodium acetate and an equal volume of ice-cold isopropanol were added to the mixture. After incubation at −20°C for 10 min, the mixture was centrifuged at 4°C at 14,000 rpm for 15 min to collect the DNA pellet, which was then washed with 1 ml ice-cold 70% (v/v) ethanol and air dried. Finally, DNA pellets were re-suspended in 200 µl AE buffer (Qiagen, Valencia, CA).
Method 5. DNA was extracted by using the DNeasy Tissue Kit (Qiagen, Valencia, CA) and the manufacturer's protocol for isolation of genomic DNA from Gram-positive bacteria was followed. Briefly, 50 µl lysozyme (10 mg/ml, Sigma-Aldrich) was added to a 500 µl aliquot of cells and the mixture was incubated for 30 min at 37°C. After the addition of 50 µl Proteinase K (20 mg/ml) and 500 µl AL buffer, the mixture was incubated for 30 min at 56°C. Then, 500 µl of ethanol was added to the lysate and the genomic DNA was purified using the columns in the kit according to the manufacturer's instructions.
Method 6. In this method, an enzymatic lysis was conducted before the PowerSoil™ DNA Isolation Kit (MO BIO Laboratories, Inc., Carlsbad, CA) was used. Briefly, 50 µl of lysozyme (10 mg/ml, Sigma-Aldrich) was added to a 500 µl aliquot of bacterial cells followed by incubation for 1 hour at 37°C. The remainder of the DNA extraction was continued beginning with step 2 of the manufacturer's protocol.
This DNA extraction experiment was finished in 12 days, in which only one DNA extraction method was used per day. The selection of DNA extraction methods was made by randomly assigning each of the six DNA extraction methods to two of 12 days. On a given day, two experimenters used a given method to extract DNA from two replicates of each sample. This was repeated once, so eight replicate samples were analyzed using each method.

Yuan S., Cohen D.B., Ravel J., Abdo Z, & Forney L.J. (2012). Evaluation of Methods for the Extraction and Purification of DNA from the Human Microbiome. PLoS ONE, 7(3), e33865.

Full text: Click here

Publication 2012

Olfactory Neuronal Calcium Imaging in Flies

Cited 50 times

Flies were reared on standard cornmeal agar medium. We used the Gal4/UAS system 41 (link) to direct the expression of the calcium sensors to PNs. GH146-Gal4 flies were a gift from L. Luo (Stanford University, Stanford, CA). UAS-GCaMP1.6 flies were a gift from D. Reiff and A. Borst (MPI, Martinsried, Germany). All experimental animals were adult females, 3–5 days after eclosion. Adult flies were dissected using previously described methods 11 (link). Flies were anaesthetized in a vial on ice just until movement stopped (<15 second) and then gently inserted into a hole in a piece of aluminum foil. Small drops of wax (55°C) were used to suspend the fly in the hole, with the edge of foil defining a horizontal plane around the head and thorax, from the first antennal segment anteriorly to the scutellum posteriorly. The dorsal side of the foil was bathed in saline, while the ventral side (including antennae and maxillary palps) remained dry and accessible to odours. A window was cut in the dorsal head cuticle between the eyes, extending from the ocelli to the first antennal segment. Fat and air sacs dorsal and anterior to the brain were removed, but the perineural sheath was left intact. The proboscis was affixed with a small drop of wax to a strand of human hair to limit brain movement. Spontaneous leg movements were typically observed in this preparation for the duration of the recording (1.5–3 hr). The saline composition used in all experiments was 42 (link) (in mM): 103 NaCl,3 KCl, 5 N-tris(hydroxymethyl) methyl-2-aminoethane-sulfonicacid, 10 trehalose, 10 glucose, 2 sucrose, 26 NaHCO₃, 1 NaH₂PO₄,1.5 CaCl₂, and 4 MgCl₂, adjusted to 275 mOsm, pH 7.3 when bubbled with 95% O₂/5%CO₂.
Odours (cis-3-hexen-1-ol (cis), and isoamyl acetate (ia)) were delivered using a custom-made odour-delivery system and a Teflon nozzle (entry diameter 1/8″) directed towards the antennae. Odours were delivered in a constant stream of air (1 l/min) at final concentrations of ca. 15%. Odour delivery times were measured using a mini-PID (Aurora Scientific Inc., Ontario, Canada). Odours were presented for either 3s or 5s. All comparisons of sensor performance were made using experiments with identical odour presentation times. The results reported are based on data obtained from 3 GCaMP1.6-expressing flies (4 ALs) and 4 GCaMP3-expressing flies (4 ALs).

Tian L., Hires S.A., Mao T., Huber D., Chiappe M.E., Chalasani S.H., Petreanu L., Akerboom J., McKinney S.A., Schreiter E.R., Bargmann C.I., Jayaraman V., Svoboda K, & Looger L.L. (2009). Imaging neural activity in worms, flies and mice with improved GCaMP calcium indicators. Nature methods, 6(12), 875-881.

Publication 2009

Olfactory Deficit Screening in Mice

Cited 18 times

Mice were screened for olfactory deficits using an odor-cross habituation test (Fig 1A)(Sundberg et al., 1982 (link); Wilson and Linster, 2008 (link)). Odors (n=7; limonene, ethyl valerate, isoamyl acetate, pentanol, heptanone, propyl butyrate and nonane; Sigma Aldrich, St. Louis, MO) were diluted 1×10⁻³ in mineral oil and applied to a cotton-applicator stick which was then enclosed in a piece of odorless plastic tubing to prevent contact of the liquid odor with the testing chamber or animal yet still allow volatile odor delivery. Notably, such an odor presentation method controls for the influence of visual and/or somatosensory influences on odor investigation. Odors were delivered for 4 successive trials (1 block), 20sec each, separated by 30sec inter-trial intervals, by inserting the odor stick into a port on the side of the animal’s home cage (Fig 1A). Home cage testing was chosen over testing in a separate apparatus to minimize potential influences of stress and anxiety (due to the new environment/context) on the behavioral measures. Testing took place during the light phase of the animals’ (12:12) day: light cycle, over two daily sessions (3–4 odors/session) separated by 24–48 hrs. The duration of time spent investigating, defined as snout-oriented sniffing within 1cm of the odor presentation port, was recorded across all trials by a single observer blind to genotypes (D.W.W.). Home cages were cleaned with fresh corn cob bedding 24–48hrs prior to behavioral testing to reduce unnecessary background odors, yet still allowing for adaptation to the new bedding. The stainless steel food bin and water bottle were removed from cages immediately prior to testing.

Wesson D.W., Levy E., Nixon R.A, & Wilson D.A. (2010). Olfactory Dysfunction Correlates with Amyloid-β Burden in an Alzheimer's Disease Mouse Model. The Journal of Neuroscience, 30(2), 505-514.

Publication 2010

Acclimatization Animals Anxiety Butyrates ethyl valerate Food Genotype Gossypium isoamyl acetate Light Limonene Maize Mice, House nonane Obstetric Delivery Odors Oil, Mineral Sense of Smell Stainless Steel Visually Impaired Persons

ChIP-exo Protocol with Detailed Steps

Cited 12 times

The main differences between our protocol and the Pugh protocol [5 (link)] are the oligonucleotides sequences, different washing buffer, the use of magnetic beads and the PCR mix. For the lambda exonuclease digestion, we have tested the Pugh conditions (10 units for 30 min) and a higher concentration (50 units for 1 h) on an ER ChIP-exo conducted in MCF-7 cells (Additional file 9). We found no significant difference in peak width with increased exonuclease concentration.
The cross-linking, cell lysis and sonication are done as described previously [17 (link)]. Each ChIP is done using 10 ug of antibody and 50 uL of Protein A or G magnetic beads (Invitrogen, Dynabeads). After the overnight ChIP on rotator at 4°C, the supernatant is removed and the beads are washed six times in 1 mL of RIPA buffer (50 mM HEPES pH 7.6; 1 mM EDTA; 0.7% Na-Deoxycholate; 1% NP-40; 0.5 M LiCL) in a 2 mL microfuge tube, followed by two washes in 1 mL of Tris HCl pH 8. The beads then undergo five successive incubations in a 2 mL tube agitated at 900 rpm in a thermomixer as followed:
1) End polishing: 1 mM ATP, 100 uM dNTP, 15 U T4 DNA polymerase, 5 U Klenow DNA polymerase, 50 U T4 PolyNucleotide Kinase, in 100 uL 1× NEBuffer 2 (50 mM NaCl, 10 mM Tris–HCl, 10 mM MgCl2, 1 mM DTT, pH 7.9) at 30°C for 30 min.
2) Ligation of the P7 exo-adapter: 1 mM ATP, 150 pmol P7 exo-adapter, 2000 U T4 DNA ligase, in 100 uL 1× NEBuffer 2 at 25°C for 60 min.
3) Nick repair: 150 uM dNTP, 15 U phi29 DNA polymerase in 100 uL 1× phi29 reaction buffer (50 mM Tris–HCl pH 7.5, 10 mM MgCl2, 10 mM (NH4)2SO4, 1 mM DTT, pH 7.5) at 30°C for 20 min.
4) Lambda exonuclease digestion: 10 U Lambda exonuclease in 100 uL 1× NEB Lambda exonuclease buffer (67 mM Glycine-KOH, 2.5 mM MgCl2, 50 μg/mL BSA, pH 9.4) at 37°C for 30 min.
5) RecJf exonuclease digestion: 30 U RecJf exonuclease in 100 uL NEBuffer 2 at 37°C for 30 min.
The beads are washed two times in 1 mL RIPA buffer and two times in 1 mL Tris HCl pH 8 after every incubation. All the incubations (1 to 5) are done so that the maximum concentration of DTT is 1 mM to avoid the elution of the ChIP material.
(6) Elution and reverse cross-linking: the beads are incubated with 100 ug of Proteinase K in 200 uL of elution buffer (50 mM Tris HCl pH 8; 10 mM EDTA; 1% SDS) overnight at 65°C. The 200 uL of supernatant is transferred to a new tube and diluted in 200 uL TE (10 mM Tris, 1 mM EDTA, pH 7.4). The DNA is purified using phenol-chloroform-isoamyl alcohol extraction followed by ethanol precipitation. The resulting DNA pellet is dissolved in 20 uL water. The DNA can be stored at this step at −20°C.
(7) P7 primer extension: the 20 uL of DNA is denaturated 5 min at 95°C, then mixed with 5 pmol of the P7 primer and incubated in 50 uL 1× NEB Phi29 reaction buffer for 5 min at 65°C and 2 min at 30°C in a thermocycler. After the addition of 10 U Phi29 DNA polymerase and 200 uM dNTP, the mix is incubated 20 min at 30°C and then 10 min at 65°C. The DNA is purified using AMPure beads (1.8 volume) and eluted in 20 uL of resuspension buffer (Tris-Acetate 10 mM pH 8).
(8) Ligation of the P5 exo-adapter: the 20 uL of DNA is mixed with 15 pmol of the P5 exo-adapter, 2,000 U T4 DNA ligase and incubated in 50 uL 1× NEB T4 DNA ligase buffer for 60 min at 25°C and then 10 min at 65°C. The DNA is purified using AMPure beads (1.8 volume) and eluted in 20 uL of resuspension buffer (Tris-Acetate 10 mM pH 8).
(9) PCR amplification: the DNA sample is amplified using 0.5 uM of the universal reverse PCR primer and the forward PCR primer containing the index sequence of choice in 50 uL 1× NEBNext High-Fidelity PCR Master Mix (New England Biolabs, M0541). The number of PCR cycles is 13 to 18, depending on the ChIP efficiency. The PCR product is purified using AMPure beads (1.8 volume) and eluted in 20 uL of resuspension buffer (Tris-Acetate 10 mM pH 8).
(10) Gel-size selection: 200 to 300 bp PCR product is purified from a 2% agarose gel using MinElute Gel Extraction Kit (Qiagen) and eluted in 20 uL of elution buffer.
(11) Illumina sequencing: the library is quantified using the KAPA library quantification kit for Illumina sequencing platforms (KAPA Biosystems, KK4824) and sequenced on a MiSeq, GAII or HiSeq following the manufacturer’s protocol.

Serandour A.A., Brown G.D., Cohen J.D, & Carroll J.S. (2013). Development of an Illumina-based ChIP-exonuclease method provides insight into FoxA1-DNA binding properties. Genome Biology, 14(12), R147.

Full text: Click here

Publication 2013

Toxicogenomic Response to Benzo[a]pyrene Exposure

Cited 11 times

Animal treatment. Adult (25 weeks old) male mice of MutaMouse strain (transgenic mouse strain 40.6) were exposed to BaP (Sigma Aldrich, Canada) as described previously (Malik et al., 2012 (link)). In brief, animals were dosed daily via oral gavage for 28 days with varying doses of BaP (0, 25, 50, and 75mg/kg body weight/day) dissolved in olive oil. Each dose group contained five animals. Mice were sacrificed by cardiac puncture under isoflorane anesthesia 72h following the final exposure. The right lobe of the lung was excised, flash frozen in liquid nitrogen, and stored at −80°C until use. For the duration of the experiment, food (2014 Teklad Global standard rodent diet) and water were provided ad libitum, and mice were caged individually in plastic film isolators (Harlan Isotec, U.K.) on a 12-h light/12-h dark cycle. Mice were bred, maintained, and treated in accordance with the Canadian Council for Animal Care Guidelines and approved by Health Canada’s Animal Care Committee.
Tissue selection. The major exposures to BaP occur via the oral route (drinking and feed) (Hettemer-Frey and Travis, 1991 (link)). The organs directly affected by BaP as a consequence of oral exposure include stomach, esophagus, tongue, and larynx (Culp et al., 1998 (link)). However, studies conducted by Stoner et al. (1984) (link) and Wattenberg and Leong (1970) (link) showed lung and liver to be equally impacted by BaP (oral gavage) and, moreover, revealed sensitivity of lung for tumor development in comparison with liver. In alignment with these reports, our previous work (Halappanavar et al., 2011 (link); Yauk et al., 2011 (link)) revealed lung-specific regulation of the biological processes known to be associated with cancer formation. Thus, in the present study, we have investigated transcriptional responses in lung and liver. The results of the study may also help address the importance of considering the multiorgan toxicity in calculating the risk associated with chemicals, such as BaP.
Tissue DNA extraction. The frozen lung and liver tissues were sliced randomly. Genomic DNA was isolated from a random tissue section for measuring the levels of DNA adducts and transgene mutant frequency. In brief, lung tissue was minced and degassed to remove all traces of air in the alveoli. Livers were homogenized in ice cold TMST buffer (50mM Tris, pH 7.6, 3mM magnesium acetate, 250mM sucrose, 0.2% Triton X-100) as described in the study by Douglas et al. (1994) (link). The minced tissue was washed twice in cold PBS and lysed in 10mM Tris, pH 7.6, 10µM EDTA, 100µM NaCl, and 1% SDS overnight at 37°C on a rotating platform. The lysate was digested with proteinase K (1mg/ml lysis buffer). DNA was isolated using a serial phenol/chloroform/isoamyl alcohol (25:24:1) and chloroform/isoamyl alcohol (24:1) extraction (Renault et al., 1997 (link)). DNA was precipitated in ethanol and dissolved in Tris-EDTA buffer (10mM Tris, pH 7.6, 500mM EDTA). DNA was stored at 4°C until use.
DNA adduct analysis. DNA adduct formation in each sample was determined using the nuclease P1 digestion enrichment version of the ³²P-postlabeling assay as described previously (Phillips and Arlt, 2007 (link)) with minor modifications. Briefly, 4 µg of DNA was digested overnight with micrococcal nuclease (288 mUnits, Sigma, cat. no. N3755) and calf spleen phosphodiesterase (1.2 mUnits, MP Biomedicals, cat. no. 100977), enriched, and labeled as described elsewhere (Phillips and Arlt, 2007 (link)). Radiolabeled adducted nucleotide biphosphates were separated by thin-layer chromatography on polyethyleneimine-cellulose plates (Macherey-Nagel, Düren, Germany) with the following chromatographic conditions (Arlt et al., 2008 (link)): D1, 1.0M sodium phosphate, pH 6; D3, 4.0M lithium formate, 7.0M urea, pH 3.5; D4, 0.8M LiCl, 0.5M Tris, 8.5M urea, pH 8. Chromatographs were scanned using a Packard Instant Imager (Canberra Packard, Downers Grove USA), and DNA adduct levels (relative adduct labeling) were calculated from the adduct counts per minute (cpm), the specific activity of [γ-³²P]ATP, and the amount of DNA (pmol of DNA-P) used. An external BPDE-DNA standard was used for identification of BaP-DNA adducts. Results are expressed as DNA adducts/10⁸ nucleotides.
LacZ mutant frequency (positive selection). The transgene lacZ mutant frequency in lungs was determined using the P-gal (phenyl-β-D-galactopyranoside) positive selection assay as described in the study by Vijg and Douglas (1996) and Lambert et al. (2005) (link). The λgt10lacZ DNA was rescued from the genomic DNA using the Transpack lambda packaging system (Stratagene). The packaged phage particles were mixed with host bacterium (Escherichia coli lacZ⁻, galE⁻, recA⁻, pAA119 with galT and galK), plated on minimal medium containing 0.3% (w/v) P-Gal, and incubated overnight at 37°C. Total plaque-forming units (pfu) were measured on concurrent titers that did not contain P-Gal. Mutant frequency is expressed as the ratio of the number of mutant pfu to total pfu. Mutant frequency data analysis was performed as described previously (Malik et al., 2012 (link)).
RNA extraction and purification. Total RNA was isolated for gene expression analysis and qRT-PCR validation as described previously (Halappanavar et al., 2011 (link)). Briefly, total RNA was extracted from the lungs using TRIzol reagent (Invitrogen) and purified using RNeasy Mini Kit (Qiagen, Canada). The RNA quantity and purity were checked using a NanoDrop Spectrophotometer (ThermoFisher Scientific, Canada). The RNA integrity was determined using an Agilent 2100 Bioanalyzer (Agilent Technologies, Canada). The samples showing A₂₆₀/A₂₈₀ ratios between 2.1 and 2.2 and having RNA integrity number above 7.5 were used for further analysis.
Microarray hybridization and analysis. Double-stranded cDNA and cyanine-labeled cRNA were synthesized (Agilent Linear Amplification Kits, Agilent Technologies) from 250ng of total RNA from each sample and universal reference total RNA (Stratagene, Canada). Cyanine-labeled cRNA targets were in vitro transcribed using T7 RNA polymerase and purified by RNeasy Mini Kit (Qiagen). From each (sample and reference) labeled sample, 825ng of cRNA was hybridized to Agilent 4×44K oligonucleotide microarrays (Agilent Technologies) at 60°C overnight (16h) in the Agilent SureHyb hybridization chamber. Arrays were washed and scanned on an Agilent G2505B Scanner according to manufacturer’s recommendations. Feature extraction software version 10.7.3.1 (Agilent Technologies) was used to extract the data.
A reference design (Kerr and Churchill, 2001 (link)) was used to analyze mRNA expression as described previously (Malik et al., 2012 (link)). All analyses were conducted in the R (R-Development-Core-Team, 2010 ) environment using the MAANOVA library (Wu, 2010 ). The background fluorescence was measured using the negative control (−)3×SLv1 probes; probes with median signal intensities less than the trimmed mean (trim = 5%) plus three trimmed standard deviations of (−)3×SLv1 probe were flagged as absent (within the background signal). Probes were considered present if at least four of the five samples within a condition had signal intensities greater than three trimmed standard deviations above the trimmed mean of the (−)3×SLv1 probes (background signal). Data were normalized using the transform.madata() function using the glowess option with a span of 0.1. Ratio intensity plots and heat maps for the raw and normalized data were constructed to identify outliers. One sample (50mg/kg group) was removed from the analysis based on clustering. The statistical model for this analysis included fixed effects of array and treatment condition and was applied to the log₂ of the absolute intensities. Differentially expressed transcripts (upregulated or downregulated relative to olive oil–treated control mouse lung samples) were determined using the Fs statistic option in the matest() function. The p values for all statistical tests were estimated by the permutation method with residual shuffling, and false discovery rate (FDR) adjusted p values were estimated using the adjPval() function. The fold change calculations were estimated as described previously (Malik et al., 2012 (link)). Significant genes were selected based on a FDR adjusted p value < 0.05 for any BaP exposed versus control contrast.
qRT-PCR array validation. Mouse pathway-specific PCR array (cancer-PAMM-033, SABiosciences) and custom PCR arrays consisting of 172 genes in total were employed to validate the microarray results. Genes for the custom array were selected based on their implication in biological processes relevant to lung carcinogenesis. These genes included statistically significant differentially expressed genes (FDR adjusted p < 0.05), differentially expressed genes that exhibited high fold changes (fold rank only) but were not statistically significant, and genes that were differentially regulated (FDR adjusted p < 0.05) in the livers from the same mice (Malik et al., 2012 (link)). In brief, 0.8 µg of total lung tissue RNA (n = 5/group) from each sample was reverse transcribed using RT² First Strand Kit (SABiosciences). Real-time PCR was performed using RT² SYBR Green PCR Master Mix on a CFX96 real-time detection system (Bio-Rad, Canada). Threshold cycle values for each well were averaged. Relative gene expression was determined according to the comparative Ct method and normalized to reference RNAs Hprt1, Gapdh, and Actb housekeeping genes for the Cancer Pathway array and to reference RNAs Gusb and Hprt1 housekeeping genes for the custom arrays. Transcripts were further normalized by subtracting the median delta Ct value for each sample. Differential expression was determined with a two-sample bootstrap test using R software (R-Development-Core-Team, 2010 ). The fold change was estimated using the ratio of the arithmetic mean of the treated sample to the mean of the control samples. Standard errors for the fold change values were estimated using the bootstrap test (Efron and Tibshirani, 1993 ).
Bioinformatics. All mRNA data are deposited in the NCBI Gene Expression Omnibus database under accession numbers GSE35718 (lung) and GSE24910 (liver). Following normalization, biological functions perturbed in response to BaP were identified using functional annotation clustering in the Database for Annotation, Visualization, and Integrated Discovery (DAVID) (Huang da et al., 2009 (link)) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) (Kanehisa and Goto, 2000 (link)). The biological and molecular functions of genes that were significantly differentially expressed (FDR adjusted p ≤ 0.05 or FDR adjusted p ≤ 0.1 and fold change ≥ 1.5) following exposure to BaP treatment were analyzed and explored in Ingenuity Pathway Analysis (IPA, Ingenuity Systems, Redwood City, CA). Molecular relational networks of genes modulated by BaP in lung tissue enriched for cancer function were generated using IPA. Each molecule was overlaid onto a global molecular network developed from information contained in the Ingenuity Knowledge Base. Networks were generated based on their connectivity. All relationships are supported by at least one reference from the literature.

Labib S., Yauk C., Williams A., Arlt V.M., Phillips D.H., White P.A, & Halappanavar S. (2012). Subchronic Oral Exposure to Benzo(a)pyrene Leads to Distinct Transcriptomic Changes in the Lungs That Are Related to Carcinogenesis. Toxicological Sciences, 129(1), 213-224.

Publication 2012

Most recents protocols related to «Isoamyl acetate»

Comprehensive Compound Library for Analytical Studies

Not available on PMC !

All compounds were purchased from Sigma-Aldrich Ltd. (Taufkirchen, Germany) and were of at least p.a. purity or higher. This included the following: methyl acetate, ethyl acetate, ethyl butyrate, octyl acetate, ethyl benzoate, propyl benzoate, hexyl hexanoate, isoamyl benzoate, isoamyl decanoate, ethyl hexanoate, 2-phenylethyl octanoate, methyl cinnamate, ethyl cinnamate, ethyl undecanoate, ethyl (E)-2-decenoate, ethyl myristate, methyl palmitate, ethyl oleate, ethyl linoleate, ethyl salicylate, diethyl succinate, methanol, ethanol, 1-propanol, isopropyl alcohol, 1-butanol, 2-butanol, isobutyl alcohol, 2-methyl butanol, isoamyl alcohol, 1-pentanol, 1hexanol, 1-heptanol, 1-octanol, 1-nonanol, 1-decanol, benzyl alcohol, 2-phenylethanol, 3-ethoxypropionaldehyde diethyl acetal, diethyl acetal, eugenol, geraniol, linalool, citronellol, α-terpineol, acetone, hexanal, heptanal, farnesene, nonanal, isovaleraldehyde, furfural, benzaldehyde, 2,4-butanedione, damascenone, and ethyl carbamate. Water purified by a Milli-Q ® Integral system supplied by Merck (Darmstadt, Germany) was used throughout the study.

Balák J., Drábová L., Schreiberová O., Doležal M., Maršík D., & Kolouchová I. (2024). Differences in Volatile Profiles and Sensory Characteristics in Plum Spirits on a Production Scale. Fermentation, 10(5), 235-235.

Publication 2024

Leaf and Callus Ultrastructure Preparation

Small parts of leaves and calluses were washed with a 0.01 M PBS buffer (pH 5.75) and fixed with 2.5% glutaraldehyde and 4% paraformaldehyde in 0.1 M of phosphate buffer, and placed under a vacuum for 24 h. After fixation the samples were dehydrated in 30, 50, 70, 80, and 90% ethanol (for 10 min each) and two times in 95% ethanol (15 min each) at room temperature. The samples were then maintained in a mixture of 95% ethanol and isoamyl acetate (1:1) for 10 min and in pure isoamyl acetate for 15 min. We used chemical drying according to Bhattacharya et al. [65 (link)]. After the isoamyl acetate, the samples were maintained in hexamethyldisilazane for 5 min at room temperature. The samples were then dried in a desiccator for 30 min and sputter-coated with palladium/gold (20/80) (coating thickness 12 nm). The samples were viewed under a Crossbeam 550 (Carl ZEISS, QEC GmbH, München, Germany) scanning electron microscope at an accelerating voltage of 3–5 kV.

Fizikova A., Subcheva E., Kozlov N., Tvorogova V., Samarina L., Lutova L, & Khlestkina E. (2024). Agrobacterium Transformation of Tea Plants (Camellia sinensis (L.) KUNTZE): A Small Experiment with Great Prospects. Plants, 13(5), 675.

Full text: Click here

Publication 2024

Scanning Electron Microscopy of Rumen-Digested Roughage

Not available on PMC !

Samples of rumen-digested roughage from the nylon bags were prepared for SEM analysis. Initially, samples were fixed in 2.5% glutaraldehyde for 24 hours. Subsequently, the samples were thoroughly rinsed with phosphate buffer and dehydrated using ethanol. This was followed by a treatment with a 1:1 mixture of isoamyl acetate and ethanol for 30 minutes, and then a subsequent treatment with pure isoamyl acetate for an hour. Post freeze-drying, the samples were mounted on specimen stubs and sputter-coated with gold. Observations and imaging were conducted using a JEOL JSM-6390A scanning electron microscope.

Ye X., Li K., Li Y., Gu M., OMOOR I.N., Liu H., Qiu S., Jiang X., Lu J., Ma Z., Wu J., & Gan Q. (2024). Deciphering the Impact of Nutrient Composition and Tissue Structure on Rumen Microbiome Dynamics in Roughage Degradation.

Publication 2024

Corneal SEM Analysis in Rabbits

Scanning electron microscopy (SEM) was performed pre-operatively and 1 day and 7 days postoperatively. The rabbit eyeballs were enucleated immediately after the rabbits were humanly killed to acquire corneal samples. After phosphate-buffered saline (PBS) rinsing, the samples were immersed in 2.5% glutaraldehyde overnight at 4°C. Then, the samples were treated with 1% Osmic acid solution, ethanol solution, and the mixture of ethanol and isoamyl acetate (V/V = 1/1), as well as pure isoamyl acetate sequentially. After drying, the sample was fixed on the metal net and was sprayed with gold in a vacuum. The image was taken by a scanning electron microscope (SU8100, HITACHI, Japan).

Liu Y., Li Y., Ji J., Fan Y., Hong J, & Wang L. (2024). A Shape Memory Polymeric Shield for Protecting Corneal Endothelium During Phacoemulsification. Translational Vision Science & Technology, 13(4), 11.

Full text: Click here

Publication 2024

Analytical Profiling of Flavour Compounds

All the dyes (Table S1) were purchased from Sigma-Aldrich (Merck KGaA, St. Louis, MI, USA). Anhydrous ethanol (≥99.5%) and ethyl acetate were purchased from Sinopharm. Trans-2-hexenal and benzaldehyde were purchased from Sigma-Aldrich. Hexyl acetate, (+)-limonene, β-myrcene, 3-carene, and isoamyl acetate were purchased from Aladdin Biochemistry Technology Co., Ltd (Shanghai, China).

Zhao M., You Z., Chen H., Wang X., Ying Y, & Wang Y. (2024). Integrated Fruit Ripeness Assessment System Based on an Artificial Olfactory Sensor and Deep Learning. Foods, 13(5), 793.

Full text: Click here

Publication 2024

Top products related to «Isoamyl acetate»

Isoamyl acetate by Merck Group

Sourced in United States, Germany, China, United Kingdom

Isoamyl acetate is a colorless, volatile liquid with a distinctive banana-like aroma. It is commonly used as a flavoring agent and in the production of various solvents and chemicals.

Su8100 by Hitachi

Sourced in Japan, United States

The SU8100 is a high-performance scanning electron microscope (SEM) manufactured by Hitachi. It is designed to provide high-resolution imaging and advanced analytical capabilities for a wide range of applications. The SU8100 features a field emission electron source, delivering excellent image quality and resolution.

Phenol chloroform isoamyl alcohol by Merck Group

Sourced in United States, Germany, United Kingdom, Australia

Phenol/chloroform/isoamyl alcohol is a solvent mixture commonly used in the extraction and purification of nucleic acids, such as DNA and RNA, from biological samples. It is effective in separating the nucleic acids from other cellular components, proteins, and organic molecules.

Hcp 2 by Hitachi

Sourced in Japan, China

The HCP-2 is a laboratory centrifuge manufactured by Hitachi. It is designed for general-purpose centrifugation applications in research and clinical laboratories. The HCP-2 features a brushless motor, digital speed and time controls, and a sturdy construction for reliable operation.

Ethyl acetate by Merck Group

Sourced in Germany, United States, India, Italy, United Kingdom, Poland, France, Spain, Sao Tome and Principe, China, Australia, Switzerland, Macao, Chile, Belgium, Brazil, Ireland, Canada, Portugal, Indonesia, Denmark, Mexico, Japan

Ethyl acetate is a clear, colorless liquid solvent commonly used in laboratory applications. It has a characteristic sweet, fruity odor. Ethyl acetate is known for its ability to dissolve a variety of organic compounds, making it a versatile tool in chemical research and analysis.

Isoamyl alcohol by Merck Group

Sourced in United States, Germany, France

Isoamyl alcohol is a colorless, flammable liquid with a characteristic, fusel-like odor. It is commonly used in laboratory settings as a solvent and reagent in various chemical processes and analyses.

Proteinase k by Thermo Fisher Scientific

Sourced in United States, Germany, Lithuania, United Kingdom, Canada, France, China, Japan, Switzerland, India, Finland, Ireland, Belgium, Portugal

Proteinase K is a highly active serine protease that hydrolyzes peptide bonds in proteins and is commonly used in molecular biology applications. It is an enzyme that digests proteins, including enzymes, to facilitate the extraction and purification of nucleic acids, such as DNA and RNA, from biological samples.

Su8010 by Hitachi

Sourced in Japan, Germany, United States, China

The SU8010 is a field emission scanning electron microscope (FE-SEM) manufactured by Hitachi. It is designed for high-resolution imaging of a wide range of sample types. The SU8010 provides stable and reliable performance, with a high-resolution capability and a wide range of analytical functions.

S 4800 by Hitachi

Sourced in Japan, United States, China, Germany, United Kingdom, Spain, Canada, Czechia

The S-4800 is a high-resolution scanning electron microscope (SEM) manufactured by Hitachi. It provides a range of imaging and analytical capabilities for various applications. The S-4800 utilizes a field emission electron gun to generate high-quality, high-resolution images of samples.

Phenol chloroform isoamyl alcohol by Thermo Fisher Scientific

Sourced in United States

Phenol/chloroform/isoamyl alcohol is a mixture commonly used in the extraction and purification of nucleic acids, such as DNA and RNA, from biological samples. It is a liquid solution that acts as a reagent in the process of isolating and separating these genetic materials from other cellular components.

More about "Isoamyl acetate"

isopentyl acetate, 3-methylbutyl acetate, SU8100, Phenol/chloroform/isoamyl alcohol, HCP-2, Ethyl acetate, Isoamyl alcohol, Proteinase K, SU8010, S-4800