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 436 can be excited with the violet laser line (405 nm) and emits at 436 nm. We recommend using a 450/50 bandpass filter, or equivalent. 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.

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: 436 nm; Laser: Violet Laser

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