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 dominant-negative function that has also been found to be upregulated in lymphoid malignancies, 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-diluted and tested by intracellular staining followed by flow cytometric analysis of mouse lymph node cells using the Foxp3/Transcription Factor Staining Buffer Set (Product # 00-5523-00) and protocol. Please refer to "Staining Intracellular Antigens for Flow Cytometry, Protocol B: One step protocol for intracellular (nuclear) proteins" located at Flow Protocols. This may be used at 5 µL (0.125 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.

Brilliant Ultra Violet™ 563 (BUV563) is a tandem dye that emits at 564 nm and is intended for use on cytometers equipped with an ultraviolet (355 nm) laser. Please make sure that your instrument is capable of detecting this fluorochrome.

When using two or more Super Bright, Brilliant Violet™, Brilliant Ultra Violet™, or other polymer dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401-42) or Brilliant Stain Buffer™ (Product # 00-4409-75) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer or Brilliant Stain Buffer for more information.

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 # 00-8222-49) (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: 350 nm; Emission: 564 nm; Laser: Ultraviolet Laser.

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