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  1. Link

    cyp2a6-antibody-ab3570.pdf

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Cardiovascular Lipids / Lipoproteins Lipid Metabolism Cytochromes
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Anti-CYP2A6 antibody (ab3570)

  • Datasheet
  • SDS
Reviews (2)Q&A (2)References (3)

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Western blot - Anti-CYP2A6 antibody (ab3570)

    Key features and details

    • Rabbit polyclonal to CYP2A6
    • Suitable for: WB
    • Reacts with: Human
    • Isotype: IgG

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    View more associated products

    Overview

    • Product name

      Anti-CYP2A6 antibody
      See all CYP2A6 primary antibodies
    • Description

      Rabbit polyclonal to CYP2A6
    • Host species

      Rabbit
    • Tested applications

      Suitable for: WBmore details
    • Species reactivity

      Reacts with: Human
    • Immunogen

      Recombinant full length protein (His-tag) corresponding to Human CYP2A6. Purified, His-tagged, full length human P450 2A6 fusion protein.

    • Positive control

      • WB: HepG2 cells.
    • General notes

      The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As

    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 or -80°C. Avoid freeze / thaw cycle.
    • Storage buffer

      Preservative: 0.05% Sodium azide
      Constituent: 99% PBS
    • Concentration information loading...
    • Purity

      IgG fraction
    • Primary antibody notes

      The Cytochrome P450 (P450) superfamily of enzymes is one of three enzyme systems which metabolize the fatty acid arachadonic acid (AA) to regulators of vascular tone. P450 enzymes are monooxygenase enzymes which require several co-factors such as nicotinamide adenine dinucleotide phosphate (NADPH) and P450 reductase. There are over 200 known genes which encode P450s. Epoxygenases are those P450s which metabolize AA to epoxyeicosatrienoic acid (EETs) and omega-hydroxylases are those P450s which produce 19- and 20-hydroxyeicosatetraenoic acids (19- and 20-HETE). As well as fatty acid metabolism, P450s also metabolize many drugs and toxins. Cytochrome P450 3A4 is abundantly expressed in liver and small intestine and is inducible by barbiturates, glucocorticoids and rifampicin.
    • Clonality

      Polyclonal
    • Isotype

      IgG
    • Research areas

      • Cardiovascular
      • Lipids / Lipoproteins
      • Lipid Metabolism
      • Cytochromes
      • Signal Transduction
      • Metabolism
      • Drug metabolism
      • Metabolism
      • Pathways and Processes
      • Metabolic signaling pathways
      • Lipid and lipoprotein metabolism
      • Lipases
      • Metabolism
      • Pathways and Processes
      • Metabolic signaling pathways
      • Drug metabolism
      • Metabolism
      • Pathways and Processes
      • Mitochondrial Metabolism
      • Cytochromes
      • Metabolism
      • Types of disease
      • Cancer

    Associated products

    • Compatible Secondaries

      • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
      • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Isotype control

      • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)

    Applications

    The Abpromise guarantee

    Our Abpromise guarantee covers the use of ab3570 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 - 1/5000. Predicted molecular weight: 57 kDa.
    Notes
    WB
    1/1000 - 1/5000. Predicted molecular weight: 57 kDa.

    Target

    • Function

      Exhibits a high coumarin 7-hydroxylase activity. Can act in the hydroxylation of the anti-cancer drugs cyclophosphamide and ifosphamide. Competent in the metabolic activation of aflatoxin B1. Constitutes the major nicotine C-oxidase. Possesses low phenacetin O-deethylation activity.
    • Tissue specificity

      Liver.
    • Sequence similarities

      Belongs to the cytochrome P450 family.
    • Cellular localization

      Endoplasmic reticulum membrane. Microsome membrane.
    • Target information above from: UniProt accession P11509 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links

      • Entrez Gene: 1548 Human
      • Omim: 122720 Human
      • SwissProt: P11509 Human
      • Unigene: 250615 Human
      • Alternative names

        • Coumarin 7 hydroxylase antibody
        • Coumarin 7-hydroxylase antibody
        • CP2A6_HUMAN antibody
        • CPA6 antibody
        • CYP2A antibody
        • CYP2A3 antibody
        • CYP2A6 antibody
        • CYPIIA6 antibody
        • Cytochrome P450 2A6 antibody
        • Cytochrome P450 IIA3 antibody
        • Cytochrome P450 subfamily IIA (phenobarbital inducible) polypeptide 6 antibody
        • Cytochrome P450(I) antibody
        • Cytochrome P450, family 2 subfamily A polypeptide 6 antibody
        • Flavoprotein linked monooxygenase antibody
        • P450C2A antibody
        • P450PB antibody
        • Xenobiotic monooxygenase antibody
        see all

      Images

      • Western blot - Anti-CYP2A6 antibody (ab3570)
        Western blot - Anti-CYP2A6 antibody (ab3570)
        All lanes : Anti-CYP2A6 antibody (ab3570) at 1/1000 dilution

        Lane 1 : Hep G2 whole cell extract
        Lane 2 : Mouse Pancreas whole cell extract

        Lysates/proteins at 30 µg per lane.

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

        Predicted band size: 57 kDa



        A 56 kDa band corresponding to Cytochrome P450 2A6 was observed in Hep G2 but not in Mouse Pancreas.

        Blocking buffer: 5 % skimmed milk.

      Protocols

      • Western blot protocols

      Click here to view the general protocols

      Datasheets and documents

      • SDS download

      • Datasheet download

        Download

      References (3)

      Publishing research using ab3570? Please let us know so that we can cite the reference in this datasheet.

      ab3570 has been referenced in 3 publications.

      • Ranjit S  et al. Extracellular Vesicles from Human Papilloma Virus-Infected Cervical Cancer Cells Enhance HIV-1 Replication in Differentiated U1 Cell Line. Viruses 12:N/A (2020). PubMed: 32098055
      • Kwon D  et al. Induction via Functional Protein Stabilization of Hepatic Cytochromes P450 upon gp78/Autocrine Motility Factor Receptor (AMFR) Ubiquitin E3-Ligase Genetic Ablation in Mice: Therapeutic and Toxicological Relevance. Mol Pharmacol 96:641-654 (2019). PubMed: 31492698
      • Deng J  et al. Circadian Regulation of Hepatic Cytochrome P450 2a5 by Peroxisome Proliferator-Activated Receptor ?. Drug Metab Dispos 46:1538-1545 (2018). PubMed: 30154104

      Customer reviews and Q&As

      Show All Reviews Q&A
      Submit a review Submit a question

      1-4 of 4 Abreviews or Q&A

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Cytochrome P450 2A6 antibody

      Excellent
      Abreviews
      Abreviews
      abreview image
      Application
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
      Sample
      Rat Tissue sections (Liver)
      Antigen retrieval step
      Heat mediated - Buffer/Enzyme Used: Citrate buffet, pH6
      Permeabilization
      No
      Specification
      Liver
      Blocking step
      BSA as blocking agent for 20 minute(s) · Concentration: 1% · Temperature: 25°C
      Fixative
      Formaldehyde
      Read More

      Abcam user community

      Verified customer

      Submitted Jul 06 2017

      Western blot abreview for Anti-CYP2A6 antibody

      Excellent
      Abreviews
      Abreviews
      abreview image
      Application
      Western blot
      Sample
      Mouse Tissue lysate - whole (liver)
      Gel Running Conditions
      Reduced Denaturing (4-20% gel)
      Loading amount
      30 µg
      Specification
      liver
      Blocking step
      Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
      Read More

      Abcam user community

      Verified customer

      Submitted Dec 13 2010

      Question

      Thank you for your response, I do not currently have enough volume to incubate my membrane with a 1:150 dilution, but I can use what I have left for about a 1:200 dilution, which I will incubate overnight tonight. I have a positive control of the Human Liver Microsomes in each blot, and have not had trouble detecting these bands, I am also looking into GAPDH and/or actin staining. Also, my adviser and I have decided to go with the Anti-Cytochrome P450 2A6 antibody (ab3570), and she has already submitted the purchase order. How would we go about receiving that discount on this item?

      Read More

      Abcam community

      Verified customer

      Asked on Apr 09 2012

      Answer

      Thank you for your response. While I cannot retroactively apply a discount to the order you have placed, the code will entitle you to xx off of one primary antibody for a future purchase. The code must be used on or before 10 October 2012. I would like to encourage you to take advantage of our Abreview program. It takes just a few minutes to leave a review and you can collect Abpoints which you may redeem for Abcam products or Amazon gift cards while at the same time sharing information about the product with your colleagues worldwide. To leave and Abreview, go to the webpage for the product, click on the Abreview Tab right below the shipping information and clickSubmit an Abreview. You can gain more Abpoints by submitting images as well. I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

      Read More

      Abcam Scientific Support

      Answered on Apr 09 2012

      Question

      Hello,

      I am a graduate student at University of the Sciences in Philadelphia, I recently ordered and received the ab53027 Anti-CYP2A6 antibody.
      I have been using it to try to detect CYP2A6 expression in both Human Liver Microsomes and microsomes I isolated from cell culture.
      I have been using it at a 1:1000 dilution - 10uL in 10mL of my blocking buffer, thus far I have had no success detecting the cell's microsomes. I have been able to consistently detect the liver microsomes so I know that my blotting procedure works, the antibody is functional and that I have an appropriate secondary antibody and chemiluminescent stain.
      On the product information insert it says to use the antibody at a 1/150 dilution for western blots, this seems quite high. Is it that the antibody is has weak affinity for CYP2A6 and needs to be at such a high concentration?
      Also, when I attempted to visit the abcam.com website for more information I found that this particular antibody is no longer available. Is this just a supply issue, or is there a functional reason for this change?
      Please advise on what volume of antibody you recommend for me to incubate with my blot, and in what volume of blocking buffer, as well as the duration of incubation i.e. 1 hour, 2 hours, overnight etc.

      Thank you for your time.

      Please let me know if you need any more clarification on my procedure.

      Stephanie L. Schaffer
      BS in Pharmacology/Toxicology 2009
      MS in Pharmacology/Toxicology 2012
      University of the Sciences in Philadelphia
      mailto:sschaffer@mail.usip.edu
      http://www.usip.edu/sga

      Read More

      Abcam community

      Verified customer

      Asked on Apr 09 2012

      Answer

      Thank you for contacting us. The recommended dilution of 1:150 is a recommendation which we make to our customers based on our experiments or those of supplier, collaborator or customer input. Although we cannot know exactly why certain products require higher dilutions in different protocols issues of affinity or reaction conditions will play a role. I would recommend using this product at the higher concentration recommended on the datasheet. I would also recommend running a positive control at the same time using the human liver Microsomes and strongly recommend using a loading control such as Beta Actin or GAPDH.They are used to normalize the levels of protein detected by confirming that protein loading is the same across the gel. The expression levels of the loading control should not vary between the different sample lanes. I have included links to these loading controls below. As you have not yet seen expression in your sample cell line I would also recommend a longer incubation, 4C overnight. We currently have a catalog of over 70,000 products. While we do make many of our products in house, we do receive others from outside suppliers. This product has been discontinued by our supplier; we do not know the exact cause for this discontinuation. We do offer a number of other CytochromeP450 2A6 antibodies which we do guarantee to work in western blot on cell lines. Our Rabbit polyclonal antibodies to Cytochrome P450 2A6, AB64890 or AB3570 may best suit your needs. As this product was discontinued shortly after your purchase I can offer you a discount off of anotherCytochrome P450 2A6 antibody should you decide to try one. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information. Beta Actin Loading Control:https://www.abcam.com/beta-Actin-antibody-mAbcam-8226-Loading-Control-ab8226.html GAPDH Loading Control:https://www.abcam.com/GAPDH-antibody-mAbcam-9484-Loading-Control-ab9484.html Further information about use of loading controls may be found at the following link:https://www.abcam.com/index.html?pageconfig=resource&rid=275 Two of our otherCytochrome P450 2A6 antibodies: AB3570:https://www.abcam.com/Cytochrome-P450-2A6-antibody-ab3570.html AB64890:https://www.abcam.com/Cytochrome-P450-2A6-antibody-ab64890.html

      Read More

      Abcam Scientific Support

      Answered on Apr 09 2012

      Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
      For licensing inquiries, please contact partnerships@abcam.com

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