Recombinant Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] (ab124921)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR6202] to Cytochrome P450 3A4/CYP3A4
- Suitable for: WB, IP, IHC-P
- Reacts with: Human
Related conjugates and formulations
Overview
-
Product name
Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202]
See all Cytochrome P450 3A4/CYP3A4 primary antibodies -
Description
Rabbit monoclonal [EPR6202] to Cytochrome P450 3A4/CYP3A4 -
Host species
Rabbit -
Specificity
Cross-reactivity with other CYP3A family: ab124921 has high possibility of reacting with CYP3A7 based on our internal test.
-
Tested applications
Suitable for: WB, IP, IHC-Pmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human Cytochrome P450 3A4/CYP3A4 aa 450-550 (C terminal). The exact sequence is proprietary.
(Peptide available asab234892) -
Positive control
- WB: Human fetal and liver cell lysates IHC-P: Human liver tissue IP: Human liver cell lysates
-
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Dissociation constant (KD)
KD = 2.63 x 10 -10 M Learn more about KD -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR6202 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
Recombinant Protein
-
Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab124921 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
---|---|---|
WB | (1) |
1/1000. Predicted molecular weight: 57 kDa.
For unpurified use at 1/10000 - 1/50000 dilution. |
IP |
1/10 - 1/100.
|
|
IHC-P |
1/400. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
For unpurified use at 1/100 - 1/250 dilution. |
Notes |
---|
WB
1/1000. Predicted molecular weight: 57 kDa. For unpurified use at 1/10000 - 1/50000 dilution. |
IP
1/10 - 1/100. |
IHC-P
1/400. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. For unpurified use at 1/100 - 1/250 dilution. |
Target
-
Function
Cytochromes P450 are a group of heme-thiolate monooxygenases. In liver microsomes, this enzyme is involved in an NADPH-dependent electron transport pathway. It performs a variety of oxidation reactions (e.g. caffeine 8-oxidation, omeprazole sulphoxidation, midazolam 1'-hydroxylation and midazolam 4-hydroxylation) of structurally unrelated compounds, including steroids, fatty acids, and xenobiotics. The enzyme also hydroxylates etoposide. -
Tissue specificity
Expressed in prostate and liver. According to some authors, it is not expressed in brain (PubMed:19094056). According to others, weak levels of expression are measured in some brain locations (PubMed:19359404 and PubMed:18545703). Also expressed in epithelium of the small intestine and large intestine, bile duct, nasal mucosa, kidney, adrenal cortex, epithelium of the gastric mucosa with intestinal metaplasia, gallbladder, intercalated ducts of the pancreas, chief cells of the parathyroid and the corpus luteum of the ovary (at protein level). -
Sequence similarities
Belongs to the cytochrome P450 family. -
Post-translational
modificationsPolyubiquitinated in the presence of AMFR and UBE2G1 and also STUB1/CHIP and UBE2D1 (in vitro). -
Cellular localization
Endoplasmic reticulum membrane. Microsome membrane. - Information by UniProt
-
Database links
- Entrez Gene: 1576 Human
- Omim: 124010 Human
- SwissProt: P08684 Human
- Unigene: 728751 Human
-
Alternative names
- 1,8-cineole 2-exo-monooxygenase antibody
- Albendazole monooxygenase antibody
- Albendazole sulfoxidase antibody
see all
Images
-
All lanes : Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] (ab124921) at 1/1000 dilution (Purified)
Lane 1 : Human liver lysates
Lane 2 : HepG2 (Human hepatocellular carcinoma epithelial cell), Whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 57 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?This antibody fails to detect BOI in normal HepG2 cells which is positive reported by PMID 26089940 and 25141173
-
Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] (ab124921) at 1/10000 dilution ((unpurified)) + Human fetal liver lysate at 10 µg
Secondary
HRP labelled goat anti rabbit at 1/2000 dilution
Predicted band size: 57 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human liver tissue sections labeling Cytochrome P450 3A4/CYP3A4 with purified ab124921 at 1/400 dilution (0.35 µg/ml). Perform heat mediated antigen retrieval using 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.
-
ab124921 (purified) at 1/20 dilution (1ug) immunoprecipitating Cytochrome P450 3A4/CYP3A4 in Human liver lysates.
Lane 1: Human liver lysates 10ug
Lane 2 (+): ab124921 & Human liver lysates
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab124921 in Human liver lysates
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was at 1/1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST. -
HepG2 cells were incubated at 37°C for 48h with vehicle control (0 μM) and different concentrations of nicardipine hydrochloride (ab120531) in DMSO. Increased expression of Cytochrome P450 3A4/CYP3A4 (ab124921, unpurified) 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 ab124921 (unpurified) at 1/10000 dilution and ab9484 at 1 μg /ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 dilution and visualised using ECL development solution.
-
ab124921 (unpurified), at a 1/100 dilution, staining Cytochrome P450 3A4/CYP3A4 in paraffin embedded Human liver tissue by Immunohistochemistry.
Protocols
Datasheets and documents
-
SDS download
-
Datasheet download
References (12)
ab124921 has been referenced in 12 publications.
- Wang X et al. Long Noncoding RNAs Hepatocyte Nuclear Factor 4A Antisense RNA 1 and Hepatocyte Nuclear Factor 1A Antisense RNA 1 Are Involved in Ritonavir-Induced Cytotoxicity in Hepatoma Cells. Drug Metab Dispos 50:704-715 (2022). PubMed: 34949673
- Zhang XB et al. Intermittent Hypoxia Inhibits Hepatic CYP1a2 Expression and Delays Aminophylline Metabolism. Evid Based Complement Alternat Med 2022:2782702 (2022). PubMed: 35529917
- Prawira A et al. Ribociclib enhances infigratinib-induced cancer cell differentiation and delays resistance in FGFR-driven hepatocellular carcinoma. Liver Int 41:608-620 (2021). PubMed: 33179425
- Wang X et al. DNA methylation determines the regulation of pregnane X receptor on CYP3A4 expression. Clin Exp Pharmacol Physiol 48:250-259 (2021). PubMed: 33048369
- Wang P et al. The Long Noncoding RNA Hepatocyte Nuclear Factor 4α Antisense RNA 1 Negatively Regulates Cytochrome P450 Enzymes in Huh7 Cells via Histone Modifications. Drug Metab Dispos 49:361-368 (2021). PubMed: 33674270