Description: The monoclonal antibody 29A1.4 recognizes mouse NKp46, also known as CD335. CD335, a member of the natural cytotoxicity receptor (NCR) family, is a glycoprotein with 2 Ig-like domains and a short cytoplasmic tail. Expression of CD335 is uniquely found on NK cells (including immature NK cells, defined as DX5- CD3-, and thereby allowing discrimination between NKT cells and NK cells (NKp46+, CD3-). Furthermore, unlike many of the NK markers which also stain NKT cells, staining with 29A1.4 is not strain specific. Staining has been shown on C57Bl/6, SJL, CBA/CA and BALB/C strains. NKp46 has been shown to play a role in NK cell-mediated lysis of several tumor cells and pathogen-infected cell lines.

The 29A1.4 monoclonal antibody has been shown to activate NK cells in vitro. The 29A1.4 monoclonal antibody does not deplete NK cells in vivo.

Applications Reported: This 29A1.4 antibody has been reported for use in flow cytometric analysis.

Applications Tested: This 29A1.4 antibody has been tested by flow cytometric analysis of mouse splenocytes. This may be used at less than or equal to 1 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.

Super Bright 645 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 645 nm. We recommend using a 660/20 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome.

When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining 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) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 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: 405 nm; Emission: 645 nm; Laser: Violet Laser

Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.