Antibody data
- Antibody Data
- Antigen structure
- References [1]
- Comments [0]
- Validations
- Flow cytometry [1]
Submit
Validation data
Reference
Comment
Report error
- Product number
- 46-9854-42 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- AHR Monoclonal Antibody (FF3399), PerCP-eFluor™ 710, 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 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.
- 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 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
No comments: Submit comment
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- 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.