BioLegend
Purified anti-human CD335 (Nkp46) Antibody
331902
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Nkp46 (CD335) is expressed by natural killer (NK cells) and has been suggested to be the most specific marker for NK cells across mammalian cell types. Our lab's focus is lymphocyte biology. Our goal in using his product was to be able to identify Nkp46-expressing cells using flow cytometry in an attempt to further study NK cell phenotypes. This particular monoclonal was selected because of its previously reported use in flow cytometry to identify NK cells (reported in the literature).
Flow Cytometry
human PBMC isolated by Ficoll
Anti-human Nkp46 antibody (BioLegend 331902), incubated with approximately 1x10^6 cells for 1 hour on ice at a 1:200 (0.25ug) or 1:400 (0.125ug) dilution in a total volume of 100ul PBS+2% FCS, 1mM EDTA, 0.1% sodium azide.
N/A
Alexa Fluor 647-conjugated donkey anti-mouse IgG (H+L) (Jackson 715 605 151), incubated for 1 hour on ice in the dark at 1:800 dilution in a total volume of 100ul PBS+2% FCS, 1mM EDTA, 0.1% sodium azide.
FACSCalibur (Becton Dickinson)
Compared to the negative staining controls (secondary ab alone and isotype control), a significant population of lymphocytes (~10%) stained positive for Nkp46 with this antibody used at a dilution of 1:200 or 1:400 measured by flow cytometry. Expression was variable in the Nkp46+ population. Nkp46- events resembled the negative controls.
For flow cytometry analysis, gating was performed on the lymphocyte population (monocytes were omitted). This product is available as a conjugated ab to a wide variety of fluorophores.
Requires little optimization. Bright staining. Well priced.
None
Highly recommended for flow cytometric identification of NK cells.