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Invitrogen
AHR Monoclonal Antibody (FF3399), PerCP-eFluor™ 710, eBioscience™
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FIGURE: 1 / 1
AHR Antibody (46-9854-42) in Flow
Intracellular staining of 3-day Th17-polarized CD4+ normal human peripheral blood cells with Anti-Human CD4 FITC (Product # 11-0049-42) and Mouse IgG2b K Isotype Control PerCP-eFluor® 710 (Product # 46-4732-82) (left) or Anti-Human AHR PerCP-eFluor® 710 (right), using the Fixation & Permeabilization Buffer Set (Product # 88-8824-00). Viable cells in the lymphocyte gate, as determined by Fixable Viability Dye eFluor® 450 (Product # 65-0863-14), were used for analysis.
Product Details
46-9854-42
Product Specifications
Species Reactivity
Human
Host/Isotype
Mouse
/ IgG2b, kappa
Recommended Isotype Control
Class
Monoclonal
Type
Antibody
Clone
FF3399
Immunogen
21H2-AHR (aa 449-848)
Conjugate
Excitation/Emission Max
482/708 nm
View spectra
Form
Liquid
Concentration
5 µL/Test
Purification
Affinity chromatography
Storage buffer
PBS, pH 7.2, with BSA
Contains
0.09% sodium azide
Storage conditions
4°C, store in dark, DO NOT FREEZE!
Shipping conditions
Ambient (domestic); Wet ice (international)
RRID
Product Specific Information
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.
Applications Tested: This FF3399 antibody has been pre-titrated and tested by intracellular staining followed by flow cytometric analysis of Th17-polarized normal human peripheral blood cells using the Intracellular Fixation & Permeabilization Buffer Set (Product # 88-8824-00) and protocol. Please refer to BestProtocols®: Protocol A: Two step protocol for (cytoplasmic) intracellular proteins located under the Resources Tab online. This can be used at 5 µL (0.06 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.
PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome.
Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-822-49) (100 µL cell sample + 100 µL IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-54) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
Excitation: 488 nm; Emission: 710 nm; Laser: Blue Laser.
Filtration: 0.2 µm post-manufacturing filtered.
Target Information
AHR (Ah Receptor) belongs to a family of proteins comprised of its dimerization partner ARNT (HIF-1 Beta) and the Drosophila proteins PER and SIM. AHR contains an N-terminal sequence of approximately 200 amino acids termed the PAS domain. AHR, found in a variety of tissues, binds to a specific DNA enhancer sequence and initiates transcription of the mRNA for the cytochrome P-450 (CYPIA1) gene. The gene for AHR encodes a ligand-activated transcription factor involved in the regulation of biological responses to planar aromatic hydrocarbons. AHR has been shown to regulate xenobiotic-metabolizing enzymes such as cytochrome P450, and its ligands included a variety of aromatic hydrocarbons. AHR is a ligand-activated helix/loop/helix transcription factor found in a variety of vertebrate species. The known ligands for AHR are foreign planar aromatic compounds, such as polycyclic aromatic compounds and halogenated aromatic compounds such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Unlike the steroid/thyroid hormone receptors, there is no known physiological ligand for AHR. Studies indicate that in non-ligand activated cells, AHR is found complexed with HSP90 predominantly in the cytoplasm. Upon binding to an agonist, the ligand-activated AhR is believed to transform to a nuclear, DNA binding form, and this transformation process appears to involve dissociation of HSP90 from AhR followed by formation of a heterodimer with AhR nuclear translocator protein (Arnt). Diseases associated with AHR include eosinophilic fasciitis and seborrheic dermatitis.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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Bioinformatics
Protein Aliases: Ah receptor; AH-receptor; aromatic hydrocarbon receptor; Aryl hydrocarbon receptor; bHLHe76; Class E basic helix-loop-helix protein 76
Gene Aliases: AHR; BHLHE76; FVH3; RP85
UniProt ID: (Human) P35869
Entrez Gene ID: (Human) 196
transcription regulatory region sequence-specific DNA binding
RNA polymerase II transcription factor activity, sequence-specific DNA binding
core promoter proximal region sequence-specific DNA binding
TFIID-class transcription factor binding
transcription coactivator binding
DNA binding
transcription factor activity, sequence-specific DNA binding
RNA polymerase II transcription factor activity, ligand-activated sequence-specific DNA binding
protein binding
TBP-class protein binding
protein homodimerization activity
protein heterodimerization activity
protein dimerization activity
Hsp90 protein binding
RNA polymerase II sequence-specific DNA binding transcription factor binding
E-box binding
sequence-specific double-stranded DNA binding
blood vessel development
regulation of adaptive immune response
negative regulation of T cell mediated immune response to tumor cell
regulation of transcription, DNA-templated
regulation of transcription from RNA polymerase II promoter
xenobiotic metabolic process
apoptotic process
response to xenobiotic stimulus
response to toxic substance
regulation of gene expression
intracellular receptor signaling pathway
regulation of B cell proliferation
circadian regulation of gene expression
negative regulation of transcription, DNA-templated
positive regulation of transcription, DNA-templated
positive regulation of transcription from RNA polymerase II promoter
rhythmic process
negative regulation of inflammatory response
intestinal epithelial structure maintenance
cellular response to molecule of bacterial origin
cellular response to cAMP
cellular response to forskolin
cellular response to 2,3,7,8-tetrachlorodibenzodioxine
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