KLRG1 Monoclonal Antibody (2F1), Super Bright™ 780, eBioscience™, Invitrogen™

Description

Description: The TER-119 monoclonal antibody reacts with mouse erythroid cells from early proerythroblast to mature erythrocyte stages. The TER-119 antigen is present in yolk sac, fetal and newborn liver, but is not expressed by cells carrying BFU-E and CFU-E activities. Several erythroleukemia cell lines tested so far are negative for expression of TER-119 antigen even after dimethylsulfoxide stimulation. Biochemical and molecular analysis of the TER-119 antigen indicate that this molecule is associated with the surface glycophorin A, but is not a typical glycophorin. Applications Reported: This TER-119 antibody has been reported for use in flow cytometric analysis. Applications Tested: This TER-119 antibody has been tested by flow cytometric analysis of mouse bone marrow cells. This may be used at less than or equal to 1.0 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 780 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 780 nm. We recommend using a 780/60 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome.

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 780 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 780 nm. We recommend using a 780/60 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 No. SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information. In some experiments, we have observed that compensation values for Super Bright 780-conjugated antibodies are higher in the violet 450/50 channel when using UltraComp eBeads microspheres (Product No. 01-2222-42) as compared to single-color stained cells. In such circumstances, we would recommend setting compensation with cells. We have also observed this in some experiments using AbC Total Antibody Compensation beads (Product No. A10497). 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 No. 00-8222) (100 μL of cell sample + 100 μL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product No. 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightnessand 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: 780 nm; Laser: Violet Laser Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company. KLRG1 (Killer cell lectin-like receptor G1, MAFA, 2F1-Ag) is a homodimeric member of the lectin-like type 2 transmembrane receptor family whose members contain characteristic immunoreceptor tyrosine-based inhibitory motifs (ITIMs) in their cytoplasmic domains. These ITIMs interact with the SH2 domains of protein phosphatases such as SHP-1. KLRG1 is an inhibitory receptor that is expressed on natural killer (NK) cells and certain T cells. NK cells are involved in the lysis of tumor cells and virus-infected cells, and mediate humoral and cell-mediated immune responses.