HELIOS Monoclonal Antibody (22F6), PerCP-eFluor™ 710, eBioscience™, Invitrogen™

Description

• Description: This 22F6 monoclonal antibody reacts with human, mouse and canine Helios
• Helios is a zinc-finger transcription factor first identified for its similarity to Ikaros
• It is expressed in hematopoietic stem cells in both the embryo and adult mice
• While the highest expression is found in regulatory T cells, Helios is not expressed in mature B cells, dendritic cells or myeloid cells
• Helios can form a homodimer as well as heterodimerize with other Ikaros family members to regulate transcription
• Eight isoforms of Helios exist, some of which possess a domit-negative function that has also been found to be upregulated in lymphoid maligcies, including T-acute lymphoblastic leukemia
• 22F6 has been reported to cross react with rat Helios
• Applications Reported: This 22F6 antibody has been reported for use in intracellular staining followed by flow cytometric analysis
• Applications Tested: This 22F6 antibody has been pre-titrated and tested by intracellular staining and flow cytometric analysis of mouse lymph node cells using the using the Foxp3/Transcription Factor Buffer Set (cat
• 00-5523) and protocol
• Please see Best Protocols Section (Staining intracellular Antigens for Flow Cytometry) for staining protocol (refer to Protocol B: One-step protocol for intracellular (nuclear) proteins)
• This can be used at 5 µL (0.00375 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 (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
• This gene encodes a member of the Ikaros family of zinc-finger proteins
• Three members of this protein family (Ikaros, Aiolos and Helios) are hematopoietic-specific transcription factors involved in the regulation of lymphocyte development
• This protein forms homo- or hetero-dimers with other Ikaros family members, and is thought to function predomitly in early hematopoietic development
• Multiple transcript variants encoding different isoforms have been found for this gene, but the biological validity of some variants has not been determined.