14-9854-82

antibody from Invitrogen Antibodies

Targeting: AHR bHLHe76

Provider product page for 14-9854-82

Western blot

Immunocytochemistry

Immunohistochemistry

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [10]
Comments [0]
Validations
Immunocytochemistry [2]
Immunohistochemistry [1]
Other assay [2]

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Product number: 14-9854-82 - Provider product page
Provider: Invitrogen Antibodies
Product name: AHR Monoclonal Antibody (FF3399), eBioscience™
Antibody type: Monoclonal
Antigen: Other
Description: Description: The FF3399 monoclonal antibody recognizes human aryl hydrocarbon receptor (AHR). The AHR is a ligand-activated transcription factor that mediates the toxic effects of a diverse group of environmental contaminants, most notably aryl hydrocarbons such as polychlorinated biphenyls (PCB) and tetrachlorodibenzo-p-dioxin (TCDD). The AHR has also been shown to bind to a number of naturally occurring compounds found in fruits and vegetables as well as compounds generated through normal cellular metabolism. AHR is localized in the cytoplasm in a complex that includes HSP90, p23, and XAP2/AIP/ARA9. Upon ligand-binding, AHR translocates to the nucleus and binds with aryl hydrocarbon receptor nuclear translocator (ARNT), and this complex binds to the consensus DNA sequence, GCGTG, found in the promoter/enhancer regions of many genes such as CYP1A1. The AHR is expressed in many cell types, with highest expression levels found in liver. The AHR has been shown to play a role in the regulation/differentiation of Treg and Th17 cells. Applications Reported: This FF3399 antibody has been reported for use in intracellular staining followed by flow cytometric analysis, western blotting, immunohistochemical staining of formalin-fixed paraffin embedded tissue sections, and immunocytochemistry (Fluorochrome-conjugated FF3399 is recommended for use in intracellular flow cytometry). Applications Tested: This FF3399 antibody has been tested by immunohistochemistry on formalin-fixed paraffin embedded tissue using high or low pH antigen retrieval solutions (cats. 00-4955 and 00-4956) by immunocytochemistry on fixed and permeabilized human cells, and by western blot. FF3399 can be used at less than or equal to 20 µg/mL. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Purity: Greater than 90%, as determined by SDS-PAGE. Aggregation: Less than 10%, as determined by HPLC. Filtration: 0.2 µm post-manufacturing filtered.
Reactivity: Human
Host: Mouse
Isotype: IgG
Antibody clone number: FF3399
Vial size: 100 µg
Concentration: 0.5 mg/mL
Storage: 4°C

AHR protein structure - 14-9854-82 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of AHR was performed using 70% confluent log phase A-375 cells. The cells were fixed with 4% Paraformaldehyde for 10 Minutes, permeabilized with 0.1% Triton™ X-100 for 10 Minutes, and blocked with 2% BSA for 10 Minutes at room temperature. The cells were labeled with AHR Monoclonal Antibody (FF3399), eBioscience (Product # 14-9854-82) at 20 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, Alexa Fluor 488 (Product # A28175), (1:2,000 dilution) for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Cytoplasmic and nuclear localization. Panel e represents MOLT-4 cells having no expression of AHR. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of AHR was performed using 70% confluent log phase A-375 cells. The cells were fixed with 4% Paraformaldehyde for 10 Minutes, permeabilized with 0.1% Triton™ X-100 for 10 Minutes, and blocked with 2% BSA for 10 Minutes at room temperature. The cells were labeled with AHR Monoclonal Antibody (FF3399), eBioscience (Product # 14-9854-82) at 20 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, Alexa Fluor 488 (Product # A28175), (1:2,000 dilution) for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Cytoplasmic and nuclear localization. Panel e represents MOLT-4 cells having no expression of AHR. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.