RecombinantRabMAb

Recombinant Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] (ab124921)

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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

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 as ab234892)

  • Positive control

    • WB: Human fetal and liver cell lysates IHC-P: Human liver tissue IP: Human liver cell lysates

  • General notes

    This product was previously labelled as Cytochrome P450 3A4

    Mouse and Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    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
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Applications

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 1/1000. 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.

See IHC antigen retrieval protocols

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
    modifications

    Polyubiquitinated 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

    • Alternative names

      • 1,8-cineole 2-exo-monooxygenase antibody
      • Albendazole monooxygenase antibody
      • Albendazole sulfoxidase antibody
      • CP33 antibody
      • CP34 antibody
      • CP3A4_HUMAN antibody
      • CYP3 antibody
      • CYP3A antibody
      • CYP3A3 antibody
      • CYP3A4 antibody
      • CYPIIIA3 antibody
      • CYPIIIA4 antibody
      • Cytochrome P450 3A3 antibody
      • Cytochrome P450 3A4 antibody
      • Cytochrome P450 family 3 subfamily A polypeptide 4 antibody
      • Cytochrome P450 HLp antibody
      • Cytochrome P450 NF-25 antibody
      • Cytochrome P450 subfamily IIIA polypeptide 4 antibody
      • cytochrome P450, subfamily IIIA (niphedipine oxidase), polypeptide 3 antibody
      • cytochrome P450, subfamily IIIA (niphedipine oxidase), polypeptide 4 antibody
      • Cytochrome P450-PCN1 antibody
      • Glucocorticoid inducible P450 antibody
      • HLP antibody
      • MGC126680 antibody
      • NF 25 antibody
      • NF25 antibody
      • Nifedipine oxidase antibody
      • P450 III steroid inducible antibody
      • P450 PCN1 antibody
      • P450, family III antibody
      • P450C3 antibody
      • P450PCN1 antibody
      • Quinine 3 monooxygenase antibody
      • Quinine 3-monooxygenase antibody
      • Taurochenodeoxycholate 6 alpha hydroxylase antibody
      • Taurochenodeoxycholate 6-alpha-hydroxylase antibody
      see all

    Images

    • Western blot - Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] (ab124921)
      Western blot - Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] (ab124921)
      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

    • Western blot - Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] (ab124921)
      Western blot - Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] (ab124921)
      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) - Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] (ab124921)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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 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.
    • Immunoprecipitation - Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] (ab124921)
      Immunoprecipitation - Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] (ab124921)

      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.

    • Western blot - Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] (ab124921)
      Western blot - 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 (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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] (ab124921)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] (ab124921)

      ab124921 (unpurified), at a 1/100 dilution, staining Cytochrome P450 3A4/CYP3A4 in paraffin embedded Human liver tissue by Immunohistochemistry.

    • OI-RD Scanning - Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] (ab124921)
      OI-RD Scanning - Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] (ab124921)
      Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD
    • Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] (ab124921)
      Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] (ab124921)

    References (7)

    Publishing research using ab124921? Please let us know so that we can cite the reference in this datasheet.

    ab124921 has been referenced in 7 publications.

    • Aravalli RN  et al. Hepatic Differentiation of Marmoset Embryonic Stem Cells and Functional Characterization of ESC-Derived Hepatocyte-Like Cells. Hepat Med 12:15-27 (2020). PubMed: 32104112
    • Huang KW  et al. Liver Activation of Hepatocellular Nuclear Factor-4a by Small Activating RNA Rescues Dyslipidemia and Improves Metabolic Profile. Mol Ther Nucleic Acids 19:361-370 (2020). PubMed: 31877412
    • Collins DP  et al. In vitro Differentiation of Human TERT-Transfected Multi-Lineage Progenitor Cells (MLPC) into Immortalized Hepatocyte-Like Cells. Hepat Med 12:79-92 (2020). PubMed: 32607015
    • Liu M  et al. Activating the pregnane X receptor by imperatorin attenuates dextran sulphate sodium-induced colitis in mice. Br J Pharmacol 175:3563-3580 (2018). PubMed: 29945292
    • Yanagi Y  et al. In vivo and ex vivo methods of growing a liver bud through tissue connection. Sci Rep 7:14085 (2017). IHC ; Rat . PubMed: 29074999
    • Chow EC  et al. Functional Integrity of the Chimeric (Humanized) Mouse Liver: Enzyme Zonation, Physiologic Spaces, and Hepatic Enzymes and Transporters. Drug Metab Dispos 44:1524-35 (2016). PubMed: 27342868
    • Higuchi Y  et al. Functional polymer-dependent 3D culture accelerates the differentiation of HepaRG cells into mature hepatocytes. Hepatol Res 46:1045-57 (2016). PubMed: 26724677

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