Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human, Mouse, Rat, Guinea Pig

Cited:

Human, Mouse, Rat

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, ELISA, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Simple Western, Immunoprecipitation, Knockdown Validated

Cited:

Western Blot, Flow Cytometry, Chemotaxis

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all AHR Primary Antibodies »

Product Specifications

Immunogen

Bacterially expressed human Aryl hydrocarbon Receptor (C-terminus). [UniProt# P35869]

Reactivity Notes

Rat reactivity reported in scientific literature (PMID: 23887904). Porcine reactivity reported from a verified customer review.

Localization

First cytoplasmic, upon binding with ligand and interaction with a HSP90, it translocates to the nucleus.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

96 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Description

Novus Biologicals Rabbit AHR Antibody - BSA Free (NB100-2289) is a polyclonal antibody validated for use in IHC, WB, ELISA, Flow, ICC/IF, Simple Western and IP. Anti-AHR Antibody: Cited in 10 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.

Scientific Data Images for AHR Antibody - BSA Free

Knockdown Validated: AHR Antibody - BSA Free [NB100-2289]

Western Blot: AHR Antibody - BSA Free [NB100-2289]

AHR-Antibody-Knockdown-Validated-NB100-2289-img0014.jpg
Immunohistochemistry: AHR Antibody - BSA Free [NB100-2289]

Immunohistochemistry: AHR Antibody - BSA Free [NB100-2289]

Immunohistochemistry: AHR Antibody [NB100-2289] - Staining of Aryl hydrocarbon Receptor in mouse prostate.
Immunocytochemistry/ Immunofluorescence: AHR Antibody - BSA Free [NB100-2289]

Immunocytochemistry/ Immunofluorescence: AHR Antibody - BSA Free [NB100-2289]

Immunocytochemistry/Immunofluorescence: AHR Antibody [NB100-2289] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.5% Triton X-100. The cells were incubated with anti-AHR at 5 ug/mL overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.
Western Blot: AHR AntibodyBSA Free [NB100-2289]

Western Blot: AHR AntibodyBSA Free [NB100-2289]

Western Blot: AHR Antibody [NB100-2289] - Detection of AhR in mouse liver cytosol using NB 100-2289.
Immunocytochemistry/ Immunofluorescence: AHR Antibody - BSA Free [NB100-2289]

Immunocytochemistry/ Immunofluorescence: AHR Antibody - BSA Free [NB100-2289]

Immunocytochemistry/Immunofluorescence: AHR Antibody [NB100-2289] - HepG2 cells were fixed for 10 minutes using 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-AHR at 1 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.
Flow Cytometry: AHR Antibody - BSA Free [NB100-2289]

Flow Cytometry: AHR Antibody - BSA Free [NB100-2289]

Flow Cytometry: AHR Antibody [NB100-2289] - An intracellular stain was performed on U937 cells with NB100-2289 (blue) and a matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).
Immunocytochemistry/ Immunofluorescence: AHR Antibody - BSA Free [NB100-2289]

Immunocytochemistry/ Immunofluorescence: AHR Antibody - BSA Free [NB100-2289]

Immunocytochemistry/Immunofluorescence: AHR Antibody [NB100-2289] - Aryl hydrocarbon Receptor antibody was tested in HeLa cells with DyLight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and Dylight 550 (red).
Flow Cytometry: AHR Antibody - BSA Free [NB100-2289]

Flow Cytometry: AHR Antibody - BSA Free [NB100-2289]

Flow Cytometry: AHR Antibody [NB100-2289] - An intracellular stain was performed on U-87 cells with AHR Antibody NB100-2289B (blue) and a matched isotype control (orange). Both antibodies were conjugated to Biotin. Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature, followed by Streptavidin - R-Phycoerythrin Protein (2012-1000, Novus Biologicals).
Simple Western: AHR AntibodyBSA Free [NB100-2289]

Simple Western: AHR AntibodyBSA Free [NB100-2289]

Simple Western: AHR Antibody [NB100-2289] - Image shows a specific band for Aryl hydrocarbon receptor in 0.5 mg/mL of HepG2 lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.
AHR Antibody - BSA Free

Western Blot: AHR Antibody - BSA Free [NB100-2289] -

Western Blot: AHR Antibody - BSA Free [NB100-2289] - Activation of AHR/Nrf2 pathway by PFT-alpha. (A) qPCR to detect mRNA level of CYP1A1, NQO1, HO1 & TRXR1 upon 20 h PFT-alpha treatment (20 µM) in MCF7 p53KO cells (upper part) & T47D cells (lower part). Relative expression level of CYP1A1 is shown in log2 scale; relative expression level of NQO1, HO1 & TRXR1 are shown as fold change, both normalized with DMSO treatment. All values represent the mean ± SD of two times independent experiments performed in three replicates. (B) Knock-down efficiency of AHR siRNA as detected by qPCR (upper part) & western blot (lower part). Relative expression level is shown as fold change normalized to scramble siRNA. All values represent the mean ± SD of two independent experiments performed in three replicates. (C) DCF-DA staining of ROS levels upon doxorubicin treatment (1 µM, 8 h) with or without PFT-alpha (20 µM, 12 h pre-treatment) in MCF7 p53KO cells transfected with scramble siRNA & siAHR. For quantification of ROS levels (lower panel), the values are reported as percentage relative to DMSO treatment group. (D) qPCR to detect mRNA level of Nrf2 targets NQO1, HO1 & TRXR1 upon 20 h PFT-alpha treatment (20 µM) in MCF7 p53KO cells transfected with scramble siRNA & siAHR. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31974452), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
AHR Antibody - BSA Free

AHR in A431 Human Cell Line.

AHR was detected in immersion fixed A431 human skin carcinoma cell line using Rabbit anti-AHR Affinity Purified Polyclonal Antibody conjugated to FITC (Catalog # NB100-2289F) (green) at 10 µg/mL overnight at 4C. Cells were counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.

Applications for AHR Antibody - BSA Free

Application
Recommended Usage

ELISA

1:100 - 1:2000

Flow Cytometry

2.5 ug/ml

Immunocytochemistry/ Immunofluorescence

1:500-1:1000

Immunohistochemistry

1:100

Immunohistochemistry-Paraffin

1:100

Immunoprecipitation

1:10-1:500

Simple Western

1:200

Western Blot

1:500-1:2000
Application Notes

In Western blot a band is seen ~90 to 105 kDa representing AHR (molecular weight varies by species and by strain). Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
See Simple Western Antibody Database for Simple Western validation: Tested in HepG2 lysate 0.5 mg/mL, separated by Size, antibody dilution of 1:200, apparent MW was 74 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.
The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Reviewed Applications

Read 2 reviews rated 4.5 using NB100-2289 in the following applications:

Flow Cytometry Panel Builder

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C. Do not freeze.

Background: AHR

Aryl hydrocarbon Receptor (AhR) is a ligand-activated transcriptional activator that binds to XRE promoter region of its target genes and has been established to activate phase I/II xenobiotic biotransformation enzyme genes. AhR binds MYBBP1A and its efficient DNA binding requires dimerization with another bHLH proteinsuch as ARNT. Besides interacting with NEDD8 and IVNS1ABP, AhR also interacts with coactivators including SRC-1, RIP140 and NOCA7, and with the corepressor SMRT. AhR localize to cytoplasm and upon binding with ligand followed by interaction with a HSP90, it translocates to the nucleus. AhR has been shown to be induced or repressed by TGFB1 as well as dioxin in a cell-type specific fashion, whereas cAMP, retinoic acid, TPA (12-O-tetradecanoyl phorbol-13 acetate) have been shown to exert repressive effects on AhR. As a member of bHLH-PAS (basic helix-loop-helix Per/ARNT/Sim) family, AhR has been implicated in physiological processes such as circadian cycle, neurogenesis, organs development and regulation of cell-cycle as well as the alternative signaling pathways (independently of exposure to xenobiotics). AhR-null mice exhibit infertility, liver abnormalities and cardiovascular problems such as a defective closure of the ductus venosus, indicative of the implication of the AhR in various developmental processes.

Long Name

Aryl Hydrocarbon Receptor

Alternate Names

BHLHE76

Gene Symbol

AHR

Additional AHR Products

Product Documents for AHR Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

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Product Specific Notices for AHR Antibody - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for AHR Antibody - BSA Free

Customer Reviews for AHR Antibody - BSA Free (2)

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Customer Images

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  • AHR Antibody
    Name: Anonymous
    Application: Immunohistochemistry-Paraffin
    Sample Tested: lung
    Species: Pig
    Verified Customer | Posted 10/26/2020
    FFPE pig lung was deparaffinized and antigen retrieval was performed. Block in 1%BSA/PBS and incubate with antibody at a dilution of 1:100 for 1 hour at room temperature. 20X image.
    AHR Antibody - BSA Free NB100-2289
  • Name: Anonymous
    Application: ELISA
    Sample Tested: Guinea Pig Liver Extract
    Species: Other
    Verified Customer | Posted 06/04/2009

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Protocols

View specific protocols for AHR Antibody - BSA Free (NB100-2289):


Immunocytochemistry Protocol

Culture cells to appropriate density on suitable glass coverslips in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and add 10% formalin to the dish. Fix at room temperature for 5-10 minutes.
2. Remove the formalin and add 0.5% Triton-X 100 in TBS to permeabilize the cells. Incubate for 5-10 minutes.
3. Remove the permeabilization buffer and add wash buffer (i.e. PBS or PBS with 0.1% Tween-20). Be sure to not let the specimen dry out. Gently wash three times for 10 minutes.
4. Alternatively, cells can be fixed with -20C methanol for 10 min at room temperature. Remove the methanol and rehydrate in PBS for 10 min before proceeding.
5. To block nonspecific antibody binding incubate in 10% normal goat serum for 1 hour at room temperature.
6. Add primary antibody at appropriate dilution and incubate at room temperature for 1 hour or at 4 degrees C overnight.
7. Remove primary antibody and replace with wash buffer. Gently wash three times for 10 minutes.
8. Add secondary antibody at the appropriate dilution. Incubate for 1 hour at room temperature.
9. Remove antibody and replace with wash buffer. Gently wash three times for 10 minutes.
10. Nuclei can be staining with 4',6' diamino phenylindole (DAPI) at 0.1 ug/ml, or coverslips can be directly mounted in media containing DAPI.
11. Cells can now be viewed with a fluorescence microscope.

*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow proper laboratory procedures for the disposal of formalin.


Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes.

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in wash buffer for 5 minutes.
3. Block each section with 100-400 ul blocking solution for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4 degrees C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature.
7. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
8. Add 100-400 ul Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
9. Wash sections three times in wash buffer for 5 minutes each.
10. Add 100-400 ul DAB substrate to each section and monitor staining closely.
11. As soon as the sections develop, immerse slides in deionized water.
12. Counterstain sections in hematoxylin.
13. Wash sections in deionized water two times for 5 minutes each.
14. Dehydrate sections.
15. Mount coverslips.


Western Blot Protocol
1. Perform SDS-PAGE on samples to be analyzed, loading 40 ug of total protein per lane.
2. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot.
5. Block the membrane using standard blocking buffer for at least 1 hour.
6. Wash the membrane in wash buffer three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature.
8. Wash the membrane in wash buffer three times for 10 minutes each.
9. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions.
Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%.

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