NK1.1 Monoclonal Antibody (PK136), PE-eFluor™ 610, eBioscience™, Invitrogen™

Description

Description: The PK136 monoclonal antibody reacts with mouse NK1.1, an antigen expressed by natural killer cells and a subset of T cells in the NK1.1 mouse strains including C57BL and NZB. Several commonly used laboratory mouse strains such as BALB/c, SJL, AKR, CBA, C3H and A do not express the NK1.1 antigen. For detection of NK cells in these strains the monoclonal antibody DX5 (Cat. No. 14-5971) should be used. Simultaneous staining of C57BL/6 spleen cells with PK136 and DX5 reveals coexpression of both markers by a majority of cells as well as presence of small populations of DX5+PK136- and DX5-PK136+ cells. Applications Reported: This PK136 antibody has been reported for use in flow cytometric analysis. Applications Tested: This PK136 antibody has been tested by flow cytometric analysis of mouse splenocytes. This can be used at less than or equal to 0.5 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. PE-eFluor™ 610 can be excited with laser lines from 488-561 nm and emits at 607 nm. We recommend using a 610/20 band pass filter (equivalent to PE-Texas Red™). Please make sure that your instrument is capable of detecting this fluorochome.

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 (cat. 00-8222) (100 µL of cell sample + 100 µL of 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: 488-561 nm; Emission: 607 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser. Filtration: 0.2 μm post-manufacturing filtered. NK1.1 (Killer cell lectin-like receptor subfamily B, member 1, KLRB1, NKR-P1A, CD161, cluster of differentiation 161), refers to Natural Killer (NK) cells, lymphocytes that mediate cytotoxicity and secrete cytokines after immune stimulation. Several genes of the C-type lectin superfamily, including the rodent NKRP1 family of glycoproteins, are expressed by NK cells and may be involved in the regulation of NK cell function. The KLRB1 protein contains an extracellular domain with several motifs characteristic of C-type lectins, a transmembrane domain, and a cytoplasmic domain. The KLRB1 protein, NKR-P1A or CD161, is classified as a type II membrane protein because it has an external C terminus. NKR-P1A, the receptor encoded by the KLRB1 gene, recognizes Lectin Like Transcript-1 (LLT1) as a functional ligand.