Overview

  • Product name
    Anti-Cytochrome P450 3A4 antibody
    See all Cytochrome P450 3A4 primary antibodies
  • Description
    Rabbit polyclonal to Cytochrome P450 3A4
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Other Immunogen Type (His-tag) corresponding to Human Cytochrome P450 3A4 . Purified, His-tagged, full length human P450 3A4 fusion protein.

  • Positive control
    • Rat liver lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab3572 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 1/2000. Detects a band of approximately 52 kDa (predicted molecular weight: 57 kDa). A 46kDa band is detected in rat liver extracts.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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

  • All lanes : Anti-Cytochrome P450 3A4 antibody (ab3572) at 1/2000 dilution

    Lane 1 : Mouse liver with Skimmed milk
    Lane 2 : Rat liver with Skimmed milk

    Lysates/proteins at 30 µg per lane.

    Blocking peptides at 5 % per lane.

    Secondary
    All lanes : HRP conjugated Goat anti-Rabbit IgG (H+L) at 1/5000 dilution

    Predicted band size: 57 kDa

  • ab3572 (1µg/ml) staining cytochrome P450 3A4 in human Liver using an automated system (DAKO Autostainer Plus). Using this protocol there is strong staining of cytoplasm of the plates of hepatic cells .
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers citrate pH 6.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
  • Western blot of cytochrome P450 3A4 on rat liver extract using ab3572.
  • ab3572 at a dilution of 1/2000 staining cytochrome P450 3A4 in rat liver extract by Western blot.
  • Anti-Cytochrome P450 3A4 antibody (ab3572) at 1/500 dilution + Human liver tissue lysate - total protein (ab29889) at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 57 kDa
    Observed band size: 52 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 26 kDa, 29 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 90 seconds

References

This product has been referenced in:
  • Chan CYS  et al. Derivation of CYP3A4 and CYP2B6 degradation rate constants in primary human hepatocytes: A siRNA-silencing-based approach. Drug Metab Pharmacokinet 33:179-187 (2018). Read more (PubMed: 29921509) »
  • Okada N  et al. Extracts of Immature Orange (Aurantii fructus immaturus) and Citrus Unshiu Peel (Citri unshiu pericarpium) Induce P-Glycoprotein and Cytochrome P450 3A4 Expression via Upregulation of Pregnane X Receptor. Front Pharmacol 8:84 (2017). Read more (PubMed: 28270768) »
See all 14 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Answer

Thank you for your enquiry. I can confirm that this product detects cytochrome P450 3A4 from human and rat tissues and cells. Unfortunately, we do not have any information or data showing cross reactivity with 3A5 but it may detect, to a lesser extent, other P450 isoforms. I am sorry there is not much information on the matter but you have any other questions, please do not hesitate to contact me.

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Question
Answer

I was informed by the originator of the antibody that the concentration was not determined, quite surprisingly. They predict the concentration to be around 1mg/ml. I'm sorry I couldn't help you more. Please do not hesitate to contact me again if I can be of further assistance,

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Answer

It is true that rats do not express 3A4. This is a human specific CYP. Rats do express others in the CYP family which this antibody cross-reacts with since the immunogen was full length human P450 3A4 fusion protein. That explains the cross-reaction with other rat CYP families (only we don't know which exact one!). If you have any more questions, please contact us again.

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Answer

Thank you very much for the details that you have provided and for your patience. What was the lot number that you received? Why did you not heat the samples to denature the proteins? In addition to the non-specific bands that you are seeing, did you see the expected band at 46 kDa? I can offer you a replacement vial or a refund, please send me the Abcam order number of purchase order number that was used.

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Question
Answer

Thank you for your enquiry. We did not perform experiments to determine the approximate concentration for these lots. It is purified IgG, and we don't know how much of it is specific for Cytochrome P450 3A4. We would recommend using this antibody at 1/2000 dilution for Western blot application.

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Answer

Thank you for your enquiry. We do not routinely offer free or trial sized samples for testing purposes. Our policy at Abcam is that if an antibody does not work as specified on the datasheet, we will offer a replacement or reimbursement. Should you decide to test an antibody in an application for which we do not have any information, please let us know how you get on and in return we will forward a GBP10/ USD15/ EUR15 Amazon gift voucher. If you have any more questions, please contact us again.

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Answer

We do not routinely offer free or trial sized samples for testing purposes. Our policy at Abcam is that if an antibody does not work as specified on the datasheet, we will offer a replacement or reimbursement. This antibody has been tested in Western Blotting and cross reacts with human and rat. If you plan to use the antibody in an application other than Western Blotting, please let us know and I will look into offering you a discount on your first vial.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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