Anti-Aryl hydrocarbon Receptor antibody [RPT1] - ChIP Grade (ab2770)

Overview

  • Product nameAnti-Aryl hydrocarbon Receptor antibody [RPT1] - ChIP Grade
    See all Aryl hydrocarbon Receptor primary antibodies
  • Description
    Mouse monoclonal [RPT1] to Aryl hydrocarbon Receptor - ChIP Grade
  • SpecificityDetects the aryl hydrocarbon receptor (AhR).
  • Tested applicationsSuitable for: IP, IHC-P, ChIP, ELISA, Flow Cyt, ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Pig
    Predicted to work with: Rabbit
  • Immunogen

    Synthetic peptide corresponding to Mouse Aryl hydrocarbon Receptor aa 12-31. Amino acids 18-21 omitted

Properties

Applications

Our Abpromise guarantee covers the use of ab2770 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
IP Use at an assay dependent concentration. Although ab2770 can be used in immunoprecipitation procedures, clone RPT9 is recommended for this procedure.
IHC-P 1/10 - 1/100. PubMed: 19617203
ChIP Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
Flow Cyt 1/100.
ICC/IF 1/100 - 1/1000.
WB 1/500 - 1/2000. Detects a band of approximately 95 kDa (predicted molecular weight: 94 kDa).

Target

  • FunctionLigand-activated transcriptional activator. Binds to the XRE promoter region of genes it activates. Activates the expression of multiple phase I and II xenobiotic chemical metabolizing enzyme genes (such as the CYP1A1 gene). Mediates biochemical and toxic effects of halogenated aromatic hydrocarbons. Involved in cell-cycle regulation. Likely to play an important role in the development and maturation of many tissues.
  • Tissue specificityExpressed in all tissues tested including blood, brain, heart, kidney, liver, lung, pancreas and skeletal muscle.
  • Sequence similaritiesContains 1 basic helix-loop-helix (bHLH) domain.
    Contains 1 PAC (PAS-associated C-terminal) domain.
    Contains 2 PAS (PER-ARNT-SIM) domains.
  • Cellular localizationCytoplasm. Nucleus. Initially cytoplasmic; upon binding with ligand and interaction with a HSP90, it translocates to the nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • Ah receptor antibody
    • AhR antibody
    • AHR_HUMAN antibody
    • Aromatic hydrocarbon receptor antibody
    • Aryl hydrocarbon receptor antibody
    • Aryl hydrocarbon receptor precursor antibody
    • bHLHe76 antibody
    • Class E basic helix loop helix protein 76 antibody
    • Class E basic helix-loop-helix protein 76 antibody
    • HGNC:348 antibody
    see all

Anti-Aryl hydrocarbon Receptor antibody [RPT1] - ChIP Grade images

  • Immunofluorescent analysis of Aryl Hydrocarbon Receptor in NIH-3T3 cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control, right) or with a Aryl Hydrocarbon Receptor monoclonal antibody (ab2770, left) at a dilution of 1:200 overnight at 4 C and incubated with a DyLight-488 conjugated secondary antibody. Aryl Hydrocarbon Receptor staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

  • Immunofluorescent analysis of Aryl Hydrocarbon Receptor in MCF-7 cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control, right) or with a Aryl Hydrocarbon Receptor monoclonal antibody (ab2770, left) at a dilution of 1:100 overnight at 4 C and incubated with a DyLight-488 conjugated secondary antibody. Aryl Hydrocarbon Receptor staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

  • Immunofluorescent analysis of Aryl Hydrocarbon Receptor in HeLa cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control, right) or with a Aryl Hydrocarbon Receptor monoclonal antibody (ab2770, left) at a dilution of 1:200 overnight at 4 C and incubated with a DyLight-488 conjugated secondary antibody. Aryl Hydrocarbon Receptor staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

  • Immunohistochemistry was performed on normal biopsies of deparaffinized Human skeletal muscle tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer and microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature and probed with a Aryl Hydrocarbon Receptor monoclonal antibody (ab2770) at a dilution of 1:20 or without primary antibody (negative control) overnight at 4�C in a humidified chamber. Tissues were washed with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
  • Immunohistochemistry was performed on cancer biopsies of deparaffinized Human breast carcinoma tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer and microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature and probed with a Aryl Hydrocarbon Receptor monoclonal antibody (ab2770) at a dilution of 1:100 or without primary antibody (negative control) overnight at 4�C in a humidified chamber. Tissues were washed with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
  • ab2770 staining aryl hydrocarbon receptor in MDA-MB-231 cells treated with nifedipine (ab120135), by ICC/IF. Increase in aryl hydrocarbon receptor expression correlates with increased concentration of nifedipine, as described in literature.
    The cells were incubated at 37°C for 6h in media containing different concentrations of ab120135 (nifedipine) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab2770 (1/100 dilution) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-mouse polyclonal antibody (ab96879) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 94 kDa


    Exposure time : 20 minutes

    MDA-MB-231 cells were incubated at 37°C for 6h with vehicle control (0 µM) and 100 µM of esomeprazole sodium (ab120500). Increased expression of aryl hydrocarbon receptor (ab2770) correlates with an increase in esomeprazole sodium concentration, as described in literature.

    Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 20µg 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 3% milk before being incubated with ab2770 at 2 µg/ml and ab8226 at 1 µg/ml overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP (ab97040) at 1/10000 dilution and visualised using ECL development solution.

  • ab2770 staining aryl hydrocarbon receptor in MDA-MB-231 cells treated with nitrendipine (ab120139), by ICC/IF. Increase in aryl hydrocarbon receptor expression correlates with increased concentration of nitrendipine, as described in literature.
    The cells were incubated at 37°C for 6h in media containing different concentrations of ab120139 (nitrendipine) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab2770 (1/100 dilution) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-mouse polyclonal antibody (ab96879) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

  • ab2770 staining aryl hydrocarbon receptor in MDA-MB-231 cells treated with nimodipine (ab120138), by ICC/IF. Increase in aryl hydrocarbon receptor expression correlates with increased concentration of nimodipine, as described in literature.
    The cells were incubated at 37°C for 6h in media containing different concentrations of ab120138 (nimodipine) in DMSO, fixed with 100% methanol for 5 minutes at -20ºC and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab2770 (1/100 dilution) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-mouse polyclonal antibody (ab96879) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

  • ICC/IF image of ab2770 stained Mcf7 cells. The cells were 4% formaldehyde fixed (10 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 (ab2770, at a 1/1000 dilution) 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.



  • Predicted band size : 94 kDa

    Image courtesy of an anonymous Abreview.

    Lane 1 (from the left): protein marker.
    Lane 2: IP sample with a rabbit anti-HSP90 antibody, un-treatment.
    Lane 3: IP sample with a rabbit anti-HSP90 antibody treated with 10ng/ml Depsipeptide for 24 hours.
    Lane 4: IP sample with a rabbit anti-HSP90 antibody treated with 10µm Benzo(a)pyrene for 24 hours.
    Lane 5 - 7: No antibody IP samples the same order as lane 2-4.
    Lane 8-10: Whole cell lysate without IP the same order as lane 2-4.

    Primary antibody ab2770 used at a 1/1000 dilution, Goat anti-mouse (IRDye® 800CW) secondary antibody used at a 1/10000 dilution.
  • Overlay histogram showing HEK293 cells stained with ab2770 (red line). The cells were fixed with 4% paraformaldehyde (10 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. The cells were then incubated with the antibody (ab2770, 1/100 dilution) 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. This antibody gave a positive signal in HEK293 cells fixed with 80% methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

References for Anti-Aryl hydrocarbon Receptor antibody [RPT1] - ChIP Grade (ab2770)

This product has been referenced in:
  • Barroso A  et al. The Aryl Hydrocarbon Receptor Modulates Production of Cytokines and Reactive Oxygen Species and Development of Myocarditis during Trypanosoma cruzi Infection. Infect Immun 84:3071-82 (2016). Read more (PubMed: 27481250) »
  • Wang C  et al. Beta-naphthoflavone (DB06732) mediates estrogen receptor-positive breast cancer cell cycle arrest through AhR-dependent regulation of PI3K/AKT and MAPK/ERK signaling. Carcinogenesis 35:703-13 (2014). Human . Read more (PubMed: 24163404) »

See all 14 Publications for this product

Product Wall

Application Western blot
Sample Human Tissue lysate - whole (Heart, left ventricle)
Loading amount 40 µg
Specification Heart, left ventricle
Gel Running Conditions Reduced Denaturing (4-12%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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Dawn Bowles

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Submitted Jan 10 2013

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Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Catfish Purified protein (Liver Tissue)
Loading amount 35 µg
Specification Liver Tissue
Treatment 75mg/kg BaP
Gel Running Conditions Reduced Denaturing (10% Acrylamide)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 24°C
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Submitted Dec 14 2011

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