Invitrogen

AHR Monoclonal Antibody (FF3399), PerCP-eFluor™ 710, eBioscience™

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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.

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产品信息

46-9854-42

应用

建议稀释比

已发表文章

流式细胞分析 (Flow)

5 µL (0.06 µg)/test

产品规格

种属反应

Human

宿主/亚型

Mouse / IgG2b, kappa

分类

Monoclonal

类型

Antibody

克隆号

FF3399

抗原

21H2-AHR (aa 449-848)

查看免疫原

偶联物

PerCP-eFluor™ 710 PerCP-eFluor™ 710 PerCP-eFluor™ 710

激发/发射光谱

482/708 nm 查看光谱

形式

Liquid

浓度

5 µL/Test

纯化类型

Affinity chromatography

保存液

PBS, pH 7.2, with 0.2% BSA

内含物

0.09% sodium azide

保存条件

4° C, store in dark, DO NOT FREEZE!

运输条件

Ambient (domestic); Wet ice (international)

RRID

AB_2573906

产品详细信息

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 Best Protocols: 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.

靶标信息

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.

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