Quercetin, isorhamnetin, miquelianin, B[a]P, 4,6-diamidino-2-phenylindole dihydrochloride (DAPI), nuclease P1, dimethyl sulfoxide (DMSO), and Triton X-100 were purchased from Sigma-Aldrich (St. Louis, MO, USA). Minimum essential medium (MEM), penicillin/streptomycin, fetal bovine serum (FBS), sodium pyruvate, and trypsin-EDTA were purchased from Welgene (Daegu, South Korea). Phosphate-buffered saline (PBS) was purchased from Biosesang (Seongnam, South Korea). Fluorescent mounting medium was purchased from Dako (Santa Clara, CA, USA). Antibodies (anti-NRF2, AhR, CYP1A1, GSTA1, CYP1A1, CYP1B1, ABCC2, ABCC3, and β-actin) and Alexa 488-conjugated anti-rabbit secondary antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA).
Cyp1a1
Manufactured by Cell Signaling Technology
Sourced in United States
CYP1A1 is a cytochrome P450 enzyme that catalyzes the oxidation of various endogenous and exogenous compounds. It is involved in the metabolism of xenobiotics and plays a role in the activation and detoxification of certain chemicals.
Automatically generated - may contain errors
Lab products found in correlation
β actin, by Cell Signaling Technology (2 mentions)
Lsm 510, by Zeiss (2 mentions)
C digit blot scanner, by LI COR (1 mentions)
Occludin, by Cell Signaling Technology (1 mentions)
Ripa extraction buffer, by Solarbio (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
E cadherin, by Cell Signaling Technology (1 mentions)
Bca protein assay kit, by Solarbio (1 mentions)
Ecl kit, by Solarbio (1 mentions)
Miquelianin, by Merck Group (1 mentions)
Fluorescent mounting medium, by Agilent Technologies (1 mentions)
Phosphate buffered saline pbs, by Biosesang (1 mentions)
Anti nrf2, by Cell Signaling Technology (1 mentions)
Minimum essential medium (mem), by Welgene (1 mentions)
Abcc2, by Cell Signaling Technology (1 mentions)
Alexa 488 conjugated anti rabbit secondary antibodies, by Cell Signaling Technology (1 mentions)
Abcc3, by Cell Signaling Technology (1 mentions)
Quercetin, by Merck Group (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Isorhamnetin, by Merck Group (1 mentions)
5 protocols using cyp1a1
1
Molecular Mechanisms of Phytochemical-Mediated AhR Modulation
2
Protein Expression Analysis Protocol
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Following required treatments, cells were lysed and proteins separated by SDS-PAGE then electroblotted onto PVDF membranes, Membranes were blocked in Tris-buffered saline (TBS) containing 5% milk and 0.1% Tween-20 (1 hr), then incubated with 1o Abs (CYP 1A1, TfR1; Cell Signaling; O/N, 4oC). Membrane incubation with 2o Abs preceeded ECL detection. Images were examined on a C-DiGit blot scanner (LI-Cor-Biosciences). Cells were seeded at a density of 2×105 in wells with coverslips and allowed 24 hrs to attach. Following 24 hrs exposure to the test compound, live images of the cells were taken using a Zeiss LSM 510 fluorescence microscope equipped with a UV laser.
3
Protein Expression Analysis in Colonic Tissue
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The colonic tissues were lysed with RIPA extraction buffer (Solarbio, China) containing a 1% protease inhibitor cocktail. Following centrifugation at 12,000 rpm for 15 min at 4°C, the supernatant was collected and the total protein concentration was quantified using a bicinchoninic acid (BCA) protein assay kit (Solarbio, China). Proteins were separated by SDS–polyacrylamide gel electrophoresis and transferred onto polyvinylidene difluoride (PVDF) membranes (Millipore, USA). The membranes were blocked with 5% skim milk for 1.5 h and incubated with primary antibodies against PXR (Abcam, USA), E-cadherin (Cell Signaling Technology, USA), Cyp1a1 (Cell Signaling Technology, USA), and occludin (Cell Signaling Technology, USA) overnight at 4°C. The membranes were washed and incubated with HRP-labeled secondary antibodies at room temperature for 2 h, followed by signal detection with an ECL Kit (Solarbio, China).
4
Immunoblotting Analysis of HepG2 Cells
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After treatment of HepG2 liver cancer cell lines with different concentrations of EO for 24 h, cells were harvested using trypsin and washed twice with ice-cold PBS. For the immunoblotting analysis, treated cells were lysed in RIPA buffer (Santa Cruz Biotech, CA, USA) for 14 min on ice, then centrifuged at 8000× g for 15 min at 4 °C. Supernatants were collected and estimated using Bradford reagent. An equal amount of protein was denatured at 92 °C for 7 min. Denatured proteins were then separated on SDS-PAGE gels and transferred to PVDF membranes. Membranes were blocked with 5% non-fat milk at room temperature for 30 min, incubated with primary antibodies (BCL-2, 1:1000; CASPASE-3, 1:2000; CYP-1A1, 1:1000; and NFκB, 1:1500) from cell-signaling technology (CST; Beverly, MA, USA) overnight at 4 °C, and then washed and incubated with horseradish peroxidase-conjugated secondary antibody at room temperature for 1 h. Protein bands were visualized by enhanced chemiluminescence (Hisense, Thermo Scientific, Waltham, MA, USA) and analyzed using a Licor analyzer. β-actin (1:2000) was used as an internal control [54 (link)].
5
Protein Expression Analysis by Western Blot
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Cells were collected after induction and cell lysate was prepared from 1 × 10 7 cells. The proteins were resolved by SDS-PAGE and transferred to polyvinylidene uoride membranes (Sigma, USA). The membrane was blocked in 5% bovine serum albumin or nonfat dry milk in Tris-buffered saline for 2 h at room temperature, then incubated with mouse primary antibodies overnight at 4℃. The membrane was washed and incubated with primary antibodies against CYP1A1 or β-actin (all from Cell Signaling Technology, Danvers, MA, USA), and horseradish peroxidase-conjugated secondary antibodies were added. Then, the bands were detected using ECL luminescent reagents (Pierce, USA). GAPDH was used as a loading control.
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