Anti-Cytochrome P450 1A2 antibody [d15 (16VII F10F12)] (ab22717)

Overview

  • Product nameAnti-Cytochrome P450 1A2 antibody [d15 (16VII F10F12)]
    See all Cytochrome P450 1A2 primary antibodies
  • Description
    Mouse monoclonal [d15 (16VII F10F12)] to Cytochrome P450 1A2
  • SpecificityThis antibody cross reacts with CYP 1A1. This antibody does not react with rat CYP 2A1, 2B1, 2B2, 2C6, 2C7, 2C11, 4A1, 4A2 and 4A3.
  • Tested applicationsSuitable for: Flow Cyt, IHC-P, ICC/IF, ELISA, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Full length protein (Rat). Rat liver cytochrome P4501A2.

Properties

Applications

Our Abpromise guarantee covers the use of ab22717 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
Flow Cyt Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IHC-P Use a concentration of 4 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF Use a concentration of 1 - 5 µg/ml.
ELISA Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 58 kDa.

Target

  • FunctionCytochromes P450 are a group of heme-thiolate monooxygenases. In liver microsomes, this enzyme is involved in an NADPH-dependent electron transport pathway. It oxidizes a variety of structurally unrelated compounds, including steroids, fatty acids, and xenobiotics. Most active in catalyzing 2-hydroxylation. Caffeine is metabolized primarily by cytochrome CYP1A2 in the liver through an initial N3-demethylation. Also acts in the metabolism of aflatoxin B1 and acetaminophen. Participates in the bioactivation of carcinogenic aromatic and heterocyclic amines. Catalizes the N-hydroxylation of heterocyclic amines and the O-deethylation of phenacetin.
  • Tissue specificityLiver.
  • Sequence similaritiesBelongs to the cytochrome P450 family.
  • Cellular localizationEndoplasmic reticulum membrane. Microsome membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • Aryl hydrocarbon hydroxylase antibody
    • CP 12 antibody
    • CP12 antibody
    • CP1A2_HUMAN antibody
    • CYP1A2 antibody
    • CYPIA2 antibody
    • Cytochrome P(3)450 antibody
    • Cytochrome P450 1A2 antibody
    • Cytochrome P450 4 antibody
    • Cytochrome P450 family 1 polypeptide 2 antibody
    • Cytochrome P450 family 1 subfamily A polypeptide 2 antibody
    • Cytochrome P450 subfamily I aromatic compound inducible polypeptide 2 antibody
    • Cytochrome P450-P3 antibody
    • Cytochrome P4501A2 antibody
    • Dioxin inducable P3 450 antibody
    • Flavoprotein linked monooxygenase antibody
    • Microsomal monooxygenase antibody
    • P(3)450 antibody
    • P3 450 antibody
    • P450 4 antibody
    • P450 form 4 antibody
    • P450 P3 antibody
    • P450(PA) antibody
    • Xenobiotic monooxygenase antibody
    see all

Anti-Cytochrome P450 1A2 antibody [d15 (16VII F10F12)] images

  • ICC/IF image of ab22717 stained MCF7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab22717, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Ab22717 staining human normal liver. Staining is localised to the cytoplasm.
    Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 antigen retrieval buffer citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes 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 we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
  • All lanes : Anti-Cytochrome P450 1A2 antibody [d15 (16VII F10F12)] (ab22717) at 1 µg/ml

    Lane 1 : Human liver tissue lysate - total protein (ab29889)
    Lane 2 : Liver (Mouse) Tissue Lysate
    Lane 3 : Liver (Rat) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Observed band size : 58 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 30 kDa,48 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 150 seconds
  • All lanes : Anti-Cytochrome P450 1A2 antibody [d15 (16VII F10F12)] (ab22717) at 1/2500 dilution

    Lane 1 : Tissue lysate prepared from murine liver microsomes
    Lane 2 : Tissue lysate prepared from murine liver microsomes
    Lane 3 : Tissue lysate prepared from murine liver microsomes
    Lane 4 : Tissue lysate prepared from murine liver microsomes

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat anti-mouse IgG(H+L)-HRP conjugate at 1/5000 dilution
    Developed using the ECL technique

    Exposure time : 1 second

    Image courtesy of an anonymous Abreview.

    See Abreview

  • Overlay histogram showing MCF7 cells stained with ab22717 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab22717, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References for Anti-Cytochrome P450 1A2 antibody [d15 (16VII F10F12)] (ab22717)

This product has been referenced in:
  • Guo Y  et al. Quantitative proteomic and functional analysis of liver mitochondria from high fat diet (HFD) diabetic mice. Mol Cell Proteomics 12:3744-58 (2013). Mouse . Read more (PubMed: 24030101) »
  • Getachew Y  et al. Susceptibility to acetaminophen (APAP) toxicity unexpectedly is decreased during acute viral hepatitis in mice. Biochem Pharmacol 79:1363-71 (2010). WB ; Human . Read more (PubMed: 20036646) »

See all 4 Publications for this product

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - other (Liver microsomes)
Loading amount 10 µg
Specification Liver microsomes
Gel Running Conditions Non-reduced Non-Denaturing (Native)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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Submitted Aug 01 2011

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