Rabbit Recombinant Monoclonal Cytochrome P450 3A4/CYP3A4 antibody. Carrier free. Suitable for IP, WB, IHC-P and reacts with Human, Transfected cell line - Human samples.
pH: 7.2 - 7.4
Constituents: PBS
IP | WB | IHC-P | |
---|---|---|---|
Human | Tested | Tested | Tested |
Transfected cell line - Human | Not recommended | Tested | Not recommended |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Transfected cell line - Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/10000 | Notes The detection of CYP3A4 recombinant protein or over -expressed protein with a C - terminal tag yielded a negative result because the immunogen of this product is located at the C - terminal of CYP3A4. |
Species Transfected cell line - Human | Dilution info 1/1000 | Notes The detection of CYP3A4 recombinant protein or over -expressed protein with a C - terminal tag yielded a negative result because the immunogen of this product is located at the C - terminal of CYP3A4. |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Transfected cell line - Human | Dilution info - | Notes - |
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A cytochrome P450 monooxygenase involved in the metabolism of sterols, steroid hormones, retinoids and fatty acids (PubMed:10681376, PubMed:11093772, PubMed:11555828, PubMed:12865317, PubMed:14559847, PubMed:15373842, PubMed:15764715, PubMed:19965576, PubMed:20702771, PubMed:21490593, PubMed:21576599). Mechanistically, uses molecular oxygen inserting one oxygen atom into a substrate, and reducing the second into a water molecule, with two electrons provided by NADPH via cytochrome P450 reductase (NADPH--hemoprotein reductase). Catalyzes the hydroxylation of carbon-hydrogen bonds (PubMed:12865317, PubMed:14559847, PubMed:15373842, PubMed:15764715, PubMed:21490593, PubMed:21576599, PubMed:2732228). Exhibits high catalytic activity for the formation of hydroxyestrogens from estrone (E1) and 17beta-estradiol (E2), namely 2-hydroxy E1 and E2, as well as D-ring hydroxylated E1 and E2 at the C-16 position (PubMed:11555828, PubMed:12865317, PubMed:14559847). Plays a role in the metabolism of androgens, particularly in oxidative deactivation of testosterone (PubMed:15373842, PubMed:15764715, PubMed:22773874, PubMed:2732228). Metabolizes testosterone to less biologically active 2beta- and 6beta-hydroxytestosterones (PubMed:15373842, PubMed:15764715, PubMed:2732228). Contributes to the formation of hydroxycholesterols (oxysterols), particularly A-ring hydroxylated cholesterol at the C-4beta position, and side chain hydroxylated cholesterol at the C-25 position, likely contributing to cholesterol degradation and bile acid biosynthesis (PubMed:21576599). Catalyzes bisallylic hydroxylation of polyunsaturated fatty acids (PUFA) (PubMed:9435160). Catalyzes the epoxidation of double bonds of PUFA with a preference for the last double bond (PubMed:19965576). Metabolizes endocannabinoid arachidonoylethanolamide (anandamide) to 8,9-, 11,12-, and 14,15-epoxyeicosatrienoic acid ethanolamides (EpETrE-EAs), potentially modulating endocannabinoid system signaling (PubMed:20702771). Plays a role in the metabolism of retinoids. Displays high catalytic activity for oxidation of all-trans-retinol to all-trans-retinal, a rate-limiting step for the biosynthesis of all-trans-retinoic acid (atRA) (PubMed:10681376). Further metabolizes atRA toward 4-hydroxyretinoate and may play a role in hepatic atRA clearance (PubMed:11093772). Responsible for oxidative metabolism of xenobiotics. Acts as a 2-exo-monooxygenase for plant lipid 1,8-cineole (eucalyptol) (PubMed:11159812). Metabolizes the majority of the administered drugs. Catalyzes sulfoxidation of the anthelmintics albendazole and fenbendazole (PubMed:10759686). Hydroxylates antimalarial drug quinine (PubMed:8968357). Acts as a 1,4-cineole 2-exo-monooxygenase (PubMed:11695850). Also involved in vitamin D catabolism and calcium homeostasis. Catalyzes the inactivation of the active hormone calcitriol (1-alpha,25-dihydroxyvitamin D(3)) (PubMed:29461981).
CYP3A3, CYP3A4, Cytochrome P450 3A4, Albendazole monooxygenase (sulfoxide-forming), Albendazole sulfoxidase, CYPIIIA3, CYPIIIA4, Cholesterol 25-hydroxylase, Cytochrome P450 3A3, Cytochrome P450 HLp, Cytochrome P450 NF-25, Cytochrome P450-PCN1, Nifedipine oxidase, Quinine 3-monooxygenase
Rabbit Recombinant Monoclonal Cytochrome P450 3A4/CYP3A4 antibody. Carrier free. Suitable for IP, WB, IHC-P and reacts with Human, Transfected cell line - Human samples.
pH: 7.2 - 7.4
Constituents: PBS
Cross-reactivity with other CYP3A family: This antibody has high posibility of reacting with CYP3A7 based on our internal test.
ab245774 is the carrier-free version of Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] ab124921.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
Cytochrome P450 3A4 also known as CYP3A4 is an enzyme that plays an important role in the metabolism of many xenobiotics and endogenous compounds. It belongs to the cytochrome P450 superfamily and is a monooxygenase with a mass of approximately 57 kDa. CYP3A4 is primarily expressed in the liver and small intestine although it can be found in other tissues at lower levels. This enzyme catalyzes the oxidation of a wide variety of structurally unrelated compounds facilitating their excretion. Proadifen is a known inhibitor of CYP3A4 often used in research to study the effects of enzyme inhibition.
CYP3A4 is involved in the metabolism of approximately half of all drugs used in clinical settings making it one of the most significant enzymes in drug metabolism. It does not usually function as part of a larger complex but acts individually on various substrates. Besides xenobiotic metabolism CYP3A4 also metabolizes steroids and other endogenous molecules. Climbazole an antifungal agent is a substrate for CYP3A4 demonstrating the enzyme's broad substrate specificity within the body's complex biochemical environment.
CYP3A4 plays a central role in the drug metabolism pathway and the synthesis and metabolism of steroids. These pathways involve other cytochrome P450 enzymes including CYP2D6 and CYP2C9 which often work alongside CYP3A4 to metabolize compounds. The pathway's coordinated activities ensure proper drug metabolism and hormone regulation. Understanding the involvement of CYP3A4 in these pathways is critical for drug development and determining drug-drug interactions.
CYP3A4 is associated with conditions such as drug-induced liver injury and variable drug responses. Alterations in CYP3A4 activity can lead to either toxic accumulations or reduced efficacy of medications. Additionally certain liver diseases can alter CYP3A4 expression further complicating treatment outcomes. The enzyme's interaction with other proteins like those of the cytochrome P450 family means that any dysfunctions can impact a wide range of metabolic processes and influence disease progression.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] ab124921 (purified) at 1/20 dilution (1ug) immunoprecipitating Cytochrome P450 3A4/CYP3A4 in Human liver lysates.
Lane 1: Human liver lysates 10ug
Lane 2 (+): Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] ab124921 & Human liver lysates
Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] ab124921 in Human liver lysates
For western blotting, VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was at 1/1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] ab124921).
All lanes: Immunoprecipitation - Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] (Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] ab124921)
Predicted band size: 57 kDa
Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] ab124921 (unpurified), at a 1/100 dilution, staining Cytochrome P450 3A4/CYP3A4 in paraffin embedded Human liver tissue by Immunohistochemistry.
This data was developed using the same antibody clone in a different buffer formulation containing Tris glycine, glycerol and sodium azide (Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] ab124921).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human liver tissue sections labeling Cytochrome P450 3A4/CYP3A4 with purified Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] ab124921 at 1/400 dilution (0.35 μg/ml). Perform heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] ab124921).
HepG2 cells were incubated at 37°C for 48h with vehicle control (0 μM) and different concentrations of nicardipine hydrochloride (Nicardipine hydrochloride, L-type Ca2+ channel antagonist ab120531) in DMSO. Increased expression of Cytochrome P450 3A4/CYP3A4 (Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] ab124921) correlates with an increase in nicardipine hydrochloride concentration, as described in literature.
Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 10μg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] ab124921 at 1/10000 dilution and Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484 at 1 μg /ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/10000 dilution and visualised using ECL development solution.
This data was developed using the same antibody clone in a different buffer formulation containing Tris glycine, glycerol and sodium azide (Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] ab124921).
All lanes: Western blot - Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] (Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] ab124921)
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 57 kDa
Exposure time: 20min
Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] ab124921, at a 1/100 dilution, staining Cytochrome P450 3A4 in paraffin embedded Human liver tissue by Immunohistochemistry. This data was developed using the same antibody clone in a different buffer formulation containing PBS#44; BSA#44; glycerol and sodium azide (Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] ab124921)
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] ab124921).
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody [EPR20018-251] ab205606 was used as a tag loading control.
The detection of CYP3A4 recombinant protein or over -expressed protein with a C - terminal tag yielded a negative result because the immunogen of this product is located at the C - terminal of CYP3A4.
All lanes: Western blot - Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] (Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] ab124921) at 1/1000 dilution
Lane 1: 293T cells transfected with an empty vector containing a Flag tag whole cell lysate at 20 µg
Lane 2: 293T cells transfected with a human CYP3A4 expression vector containing a N-terminal Flag tag whole cell lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 50 kDa
Exposure time: 1s
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