Description: The V3GYWCH monoclonal antibody reacts with human and mouse interferon regulatory factor 8 (IRF8; ICSBP). IRF8 is a 50 kDa transcription factor that plays a critical role in the development of dendritic cells (DC). Specifically, IRF8 is essential in the development and differentiation of plasmacytoid DC (pDC) and CD8a+ DC. IRF8-/- mice are deficient in both pDC and CD8a+ DC populations. CD8a- DC are present in normal numbers in IRF8-/- mice but fail to mature upon Toll-like receptor signaling and are functionally impaired in response to in vivo microbial stimulation. IRF8 deficiency does not affect the frequency or viability of DC precursors and as a result retroviral IRF8 transduction restores pDC development from DC progenitors in IRF8-/- mice.
Applications Reported: This V3GYWCH antibody has been reported for use in intracellular staining followed by flow cytometric analysis.
Applications Tested: This V3GYWCH antibody has been tested by intracellular staining and flow cytometric analysis of normal human peripheral blood cells using the Foxp3/Transcription Factor Staining Buffer Set (cat. 00-5523) and protocol. Please refer to Best Protocols: Protocol B: One-step protocol: intracellular (nuclear) proteins located under the Resources Tab online. This can be used at less than or equal to 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. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
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 (cat. 00-8222) (100 µL cell sample + 100 µL IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) 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.
IRF8 (interferon regulatory factor 8) is a transcriptional activator or repressor. It specifically binds to the upstream regulatory region of type I IFN and IFN-inducible MHC class I genes. IRF8 plays a negative regulatory role in cells of the immune system. It is involved in CD8+ dendritic cell differentiation by forming a complex with the BATF-JUNB heterodimer in immune cells. This leads to recognition of AICE sequence, an immune-specific regulatory element, followed by cooperative binding of BATF and IRF8 and activation of the genes. Mutations in the gene can result in immunodeficiencey 32A and 32B.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Protein Aliases: H-ICSBP; ICSBP; ICSBP1; interferon consensus sequence binding protein 1; Interferon consensus sequence-binding protein; Interferon regulatory factor 8; IRF-8
Gene Aliases: AI893568; H-ICSBP; ICSBP; ICSBP1; IMD32A; IMD32B; IRF-8; IRF8; Myls