Anti-Cytochrome P450 3A4/CYP3A4 抗体 (ab3572)

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Key features and details

  • Rabbit polyclonal to Cytochrome P450 3A4/CYP3A4
  • Suitable for: WB, IHC-P
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG

製品の概要

  • 製品名

    Anti-Cytochrome P450 3A4/CYP3A4 antibody
    Cytochrome P450 3A4/CYP3A4 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to Cytochrome P450 3A4/CYP3A4

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: WB, IHC-Pmore details

  • 種交差性

    交差種: Mouse, Rat, Human

  • 免疫原

    Other Immunogen Type (His-tag) corresponding to Human Cytochrome P450 3A4/CYP3A4. Purified, His-tagged, full length human P450 3A4 fusion protein.

  • ポジティブ･コントロール

    • Rat liver lysate.

  • 特記事項

     This product was previously labelled as Cytochrome P450 3A4

     

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

ターゲット情報

  • 機能

    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.

  • 組織特異性

    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).

  • 配列類似性

    Belongs to the cytochrome P450 family.

  • 翻訳後修飾

    Polyubiquitinated in the presence of AMFR and UBE2G1 and also STUB1/CHIP and UBE2D1 (in vitro).

  • 細胞内局在

    Endoplasmic reticulum membrane. Microsome membrane.
  • Information by UniProt

  • 参照データベース

    • 別名

      • 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

    画像

    • Western blot - Anti-Cytochrome P450 3A4/CYP3A4 antibody (ab3572)
      Western blot - Anti-Cytochrome P450 3A4/CYP3A4 antibody (ab3572)
      All lanes : Anti-Cytochrome P450 3A4/CYP3A4 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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytochrome P450 3A4/CYP3A4 antibody (ab3572)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytochrome P450 3A4/CYP3A4 antibody (ab3572)

      ab3572 (1µg/ml) staining Cytochrome P450 3A4/CYP3A4 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 - Anti-Cytochrome P450 3A4/CYP3A4 antibody (ab3572)
      Western blot - Anti-Cytochrome P450 3A4/CYP3A4 antibody (ab3572)

      Western blot of Cytochrome P450 3A4/CYP3A4 on rat liver extract using ab3572.

    • Western blot - Anti-Cytochrome P450 3A4/CYP3A4 antibody (ab3572)
      Western blot - Anti-Cytochrome P450 3A4/CYP3A4 antibody (ab3572)

      ab3572 at a dilution of 1/2000 staining Cytochrome P450 3A4/CYP3A4 in rat liver extract by Western blot.

    • Western blot - Anti-Cytochrome P450 3A4/CYP3A4 antibody (ab3572)
      Western blot - Anti-Cytochrome P450 3A4/CYP3A4 antibody (ab3572)
      Anti-Cytochrome P450 3A4/CYP3A4 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

    参考文献 (21)

    ab3572 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab3572 は 21 報の論文で使用されています。

    • Reed L  et al. Application of hepatic cytochrome b5/P450 reductase null (HBRN) mice to study the role of cytochrome b5 in the cytochrome P450-mediated bioactivation of the anticancer drug ellipticine. Toxicol Appl Pharmacol 366:64-74 (2019). PubMed: 30685480
    • Zhou XY  et al. Enzymatic Activities of CYP3A4 Allelic Variants on Quinine 3-Hydroxylation In Vitro. Front Pharmacol 10:591 (2019). PubMed: 31214030
    • Katsuda T  et al. Generation of human hepatic progenitor cells with regenerative and metabolic capacities from primary hepatocytes. Elife 8:N/A (2019). PubMed: 31393263
    • Lin Y  et al. Bmal1 regulates circadian expression of cytochrome P450 3a11 and drug metabolism in mice. Commun Biol 2:378 (2019). PubMed: 31633069
    • He Z  et al. Chaihu-Shugan-San Reinforces CYP3A4 Expression via Pregnane X Receptor in Depressive Treatment of Liver-Qi Stagnation Syndrome. Evid Based Complement Alternat Med 2019:9781675 (2019). PubMed: 31781287
    View all Publications for this product

    レビューと Q&A

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    1-7 of 7 Abreviews or Q&A

    Question

    Hi, I have a question about ab3572. I performed western blot with it and got two bands which have similar size. I suppose that these are CYP3A4 and 3A5. There was no comment about cross-reactivity with CYP3A5 in the datasheet. Does ab3572 have crosse-reactivity with CYP3A5? Even if there's no data about cross-reactivity, can we expect it? I am looking forward to your kind reply. Thanks.

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    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

    What is the concentration of the current lot of ab3572?

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    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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    Question

    Customer has said that there is no 3A4 present in rat - do you have any information regarding this?

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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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    Question

    1. more bands 2. rat liver whole cell lysates 3. 50 micro grams 4. i used three different protease inhibitors and used sonication 5. no i didnt heat the samples 6. primary antibody raised in rabbit 7. 1 in 2000 8. incubated over night. did five to six washes 9. the secondary antibody concentrations were tested from 1 in 2000 to 1 in 10,000. 10. no it is not due to the secondary antibody as they look were no bands in secondary antibody control 11. incubated for one hour in secondary antibody 12. we are using horse raddish peroxidase and taking the pictures in x ray film 13. yes i did a column as control in which i didnt add the primary antibody 14. i tried different concentrations of secondary

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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

    Customer would like to know the concentration of the batch currently in stock.

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    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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    Question

    I am working on fescue toxin in rats liver. I am plannig for immuno histo chemistry in around 72 samples. I was strongly suggested to use the antibodies from ur company. so i went to abcam website. in that i saw that this antibody is tested only for WB. so i am not sure if i can use this for IHC. so i request you to send me a very small quantity of the CYTOCHROME P 450 3A4 (AB3572) sample. so that i can test run for a couple of samples.

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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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    Question

    I am writing to you with reference to your antibody to Cytochrome P450 3A4 (ab3572). I need to screen quite large numbers of transgenic cyp3A4 regulatable cells that I am developing and consequently will need an antibody to cyp3A4. Before I purchase the antibody, I will need to know if it is applicable to my research use. Therefore would it be possible to send me a very small quantity (5 or 10microL) of your anti cyp3A4 for me to test prior to purchase? Many thanks for your time.

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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. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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