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Description: This 2F1 monoclonal antibody reacts with the mouse Killer cell Lectin-like Receptor G1 (KLRG1), also known as Mast cell Function-associated Antigen (MAFA). KLRG1 is a homodimer of glycosylated 30-38 kDa subunits and contains a cytoplasmic motif similar to the immunoreceptor tyrosine-based inhibitory motif (ITIM). Rat MAFA was identified as an antigen specific to rat mast cells; however, the expression of mouse KLRG1/MAFA using 2F1 has not been detected on the surface of mouse mast cell lines, bone marrow-derived mast cells, or peritoneal mast cells. This antigen is expressed on approximately one-third of mouse NK cells and a subset of T cells. MHC class I molecules regulate KLRG1 via interactions with class I-specific inhibitory Ly49 molecules and SHP-1 signaling. Although KLRG1 and Ly49 are both lectin-like inhibitory receptors that are regulated by class I MHC expression, the effects of this on cell surface expression of these molecules are opposing, and the underlying regulatory mechanisms distinct.
Applications Reported: This 2F1 antibody has been reported for use in flow cytometric analysis.
Applications Tested: This 2F1 antibody has been tested by flow cytometric analysis of mouse splenocytes. This can 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.
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.
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 # 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 # 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 # 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: 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.
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蛋白别名: Killer cell lectin-like receptor subfamily G member 1; Mast cell function-associated antigen 2F1; natural killer cell receptor
基因别名: 2F1-Ag; Klrg1; MAFA; MAFA-L
UniProt ID: (Mouse) O88713
Entrez Gene ID: (Mouse) 50928