Antibody data
- Antibody Data
- Antigen structure
- References [2]
- Comments [0]
- Validations
- Flow cytometry [1]
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Validation data
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- Product number
- 25-9854-42 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- AHR Monoclonal Antibody (FF3399), PE-Cyanine7, 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. 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 antibody can be used at 5 µL (0.5 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. Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light. Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-822-49) (100 µL of cell sample + 100 µL of 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: 488-561 nm; Emission: 775 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser. Filtration: 0.2 µm post-manufacturing filtered.
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- FF3399
- Vial size
- 100 Tests
- Concentration
- 5 µL/Test
- Storage
- 4° C, store in dark, DO NOT FREEZE!
Submitted references Bilirubin suppresses Th17 immunity in colitis by upregulating CD39.
Generation of IL-8 and IL-9 producing CD4⁺ T cells is affected by Th17 polarizing conditions and AHR ligands.
Longhi MS, Vuerich M, Kalbasi A, Kenison JE, Yeste A, Csizmadia E, Vaughn B, Feldbrugge L, Mitsuhashi S, Wegiel B, Otterbein L, Moss A, Quintana FJ, Robson SC
JCI insight 2017 May 4;2(9)
JCI insight 2017 May 4;2(9)
Generation of IL-8 and IL-9 producing CD4⁺ T cells is affected by Th17 polarizing conditions and AHR ligands.
Gasch M, Goroll T, Bauer M, Hinz D, Schütze N, Polte T, Kesper D, Simon JC, Hackermüller J, Lehmann I, Herberth G
Mediators of inflammation 2014;2014:182549
Mediators of inflammation 2014;2014:182549
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Supportive validation
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- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- CD4+ cells were enriched from normal human peripheral blood using the MagniSort™ Human CD4 T cell Enrichment Kit (Product # 8804-6811-74) then polarized under Th17 conditions for 3 days. Cells were intracellularly stained with Anti-Human CD4 PE (Product # 12-0049-42) and Mouse IgG2b K Isotype Control PE-Cyanine7 (Product # 25-4732-81) (left) or Anti-Human AHR PE-Cyanine7 (right) using the Intracellular Fixation & Permeabilization Buffer Set (Product # 88-8824-00) and protocol. Viable cells in the lymphocyte gate, as determined by Fixable Viability Dye eFluor® 780 (Product # 65-0865-14), were used for analysis.