Cyp1a2

Manufactured by Abcam

Sourced in China

CYP1A2 is an enzyme that plays a role in the metabolism of various compounds, including certain drugs and environmental pollutants. It is a member of the cytochrome P450 enzyme family.

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Lab products found in correlation

Cyp2e1, by Abcam (2 mentions) Tolbutamide, by LGC (1 mentions) 4 hydroxytolbutamide, by LGC (1 mentions) 6β hydroxytestosterone, by LGC (1 mentions) Omeprazole, by LGC (1 mentions) Reverse transcription kit, by Tsingke (1 mentions) Testosterone, by Solarbio (1 mentions) Dextromethorphan, by LGC (1 mentions) Nadph, by Solarbio (1 mentions) 5 hydroxy omeprazole, by LGC (1 mentions) Nitrocellulose membrane, by GE Healthcare (1 mentions) Laemmli sample buffer, by Merck Group (1 mentions) Cyp2d6, by BD (1 mentions) Mammalian proteasearrest, by G Biosciences (1 mentions) Ripa lysis buffer, by Thermo Fisher Scientific (1 mentions) Goat anti rabbit igg h l, by Abcam (1 mentions) β actin, by Merck Group (1 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (1 mentions) Supersignal west pico chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions) Cyp3a4, by Abcam (1 mentions)

Spelling variants of this product

Anti-Cyp1a2 (2 citations) Goat anti-CYP1A2 (2 citations)

4 protocols using cyp1a2

Investigating Cytochrome P450 Enzymes

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The following are used in the study: testosterone, paracetamol, phenacetin, and NADPH (Beijing Solarbio Science & Technology Co., Ltd. China); dextran, dextromethorphan, omeprazole, 5-hydroxy omeprazole, tolbutamide, 4-hydroxytolbutamide, and 6β-hydroxytestosterone (Toronto Research Chemicals); BCA protein Kit (Lianke Biotechnology Corporate Limited, No. a81911151); primer (synthesized by Tsingke Biotechnology Co., Ltd.); reverse transcription Kit (Tsingke Biotechnology Co., Ltd., No. 20190328); and CYP1A2, cyp2d1, CYP3A4, CYP3A1, CYP2C19, and CYP2C11 antibodies (Abcam Company).

Tang Y, & Li Y. (2022). Compound Shuganjieyu Capsule on CYP450 Enzyme Activity. BioMed Research International, 2022, 3228738.

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Western Blot Validation of iTRAQ Results

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Samples collected in iTRAQ buffer were quantified by Bradford assay and assessed by western blot to validate iTRAQ results. Briefly, 5 µg samples were denatured at 80 °C in Laemmli sample buffer (Sigma-Aldrich, St. Louis, MO) and separated in 10 % polyacrylamide gels and then transferred to nitrocellulose membranes (G.E Healthcare, Buckinghamshire, UK). Following 1 h blocking in 10 % milk (Bio-Rad, Hercules, CA), primary antibodies directed against CYP2E1 (Abcam, Cambridge, UK; 1:5000), CYP2D6 (BD Gentest, San Jose, CA; 1:1000), CYP1A2 (Abcam, Cambridge, UK; 1:3000) were added overnight or for 15 min Actin (Abcam, Cambridge, UK; 1:10,000). Following washing, secondary mouse (1:10,000; CYP2D6, CYP1A2, Actin) or rabbit (1:5000; CYP2E1) antibodies were subsequently added for 1 h. Membranes were then washed and visualised using chemiluminescence.

Heslop J.A., Rowe C., Walsh J., Sison-Young R., Jenkins R., Kamalian L., Kia R., Hay D., Jones R.P., Malik H.Z., Fenwick S., Chadwick A.E., Mills J., Kitteringham N.R., Goldring C.E, & Kevin Park B. (2016). Mechanistic evaluation of primary human hepatocyte culture using global proteomic analysis reveals a selective dedifferentiation profile. Archives of Toxicology, 91(1), 439-452.

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Characterization of CYP Enzymes

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After transfections, cells were lysed with ice-cold RIPA lysis buffer (ThermoFisher Scientific, Carlsbad, CA) containing protease inhibitors (Pierce Protease Inhibitor Mini Tablets, Rockford, IL and Mammalian Protease Arrest, GBiosciences, St. Louis, MO) and centrifuged at 12,500 rpm for 10 min at 4 °C. Cell supernatants were used for immunoblots. Proteins were quantified using Pierce BCA Protein Assay Kit (ThermoFisher Scientific, Carlsbad, CA) and 20 ug of protein was loaded on to precast polyacrylamide Bis-tris 10% gels (GenScript, Piscataway, NJ). The membranes were blocked with 5% skim milk and incubated with primary antibodies against CYP2E1, CYP1A2 and CYP3A4 (Abcam, Cambridge, MA) and β-actin (Sigma, St. Louis, MO). Next membranes were incubated with HRP-conjugated secondary antibodies to goat anti-rabbit IgG H&L (Abcam, Cambridge, MA). The protein bands were visualized using the SuperSignal West Pico Chemiluminescent Substrate (ThermoFisher Scientific, Carlsbad, CA) and densitometry was performed with the ImageJ software program (NIH, Bethesda, MD).

Gill P., Bhattacharyya S., McCullough S., Letzig L., Mishra P.J., Luo C., Dweep H, & James L. (2017). MicroRNA regulation of CYP 1A2, CYP3A4 and CYP2E1 expression in acetaminophen toxicity. Scientific Reports, 7, 12331.

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Liver Protein Extraction and Quantification

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Three hundred milligrams of liver tissue was homogenized in NP40 lysis buffer (20 mM Tris–HCl, 137 mM NaCl, 10% glycerol, 1% non-idet P-40, and 2 mM ethylenediaminetetraacetic acid EDTA with one protease inhibitor cocktail tablet (Roche, Indianapolis, IN) per 25 mL) on the ice. Homogenized tissue was then centrifuged at 10,000×g for 30 min, and then the supernatant was transferred to a new collection vial. Protein concentrations in the collected supernatant were quantified by the Pierce BCA Protein Quantitation Assay kit (Thermo Fisher Scientific, Waltham, MA) following the manufacture’s protocol.
Fifty micrograms of the whole cell lysate in each well was separated on 7.5% SDS–PAGE gels and transferred to polyvinylidene difluoride membranes. The following antibodies were used for the detection of proteins: Erk2 (Santa Cruz Biotechnology, Dallas, TX, sc-154), Cyp1a2 (sc-9835), Cyp3a (sc-25845), Cyp2d22 (Abcam, Cambridge, MA, Ab62204), and Cyp2c29 (Ab137015). Densitometry analysis of the Western blot was performed using ImageJ software (National Institutes of Health, Bethesda, MD), and relative protein expression was calculated by normalizing to the housekeeping protein Erk2 (sc-154).

Li H., Clarke J.D., Dzierlenga A.L., Bear J., Goedken M.J, & Cherrington N.J. (2016). In vivo cytochrome P450 activity alterations in diabetic nonalcoholic steatohepatitis mice. Journal of biochemical and molecular toxicology, 31(2), 10.1002/jbt.21840.

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