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Multicolor flow cytometric analysis of KLRG1 expression on human peripheral blood lymphocytes. Human whole blood was stained with PE Mouse Anti-Human CD56 (Cat. No. 563238) and either BD Horizon™ BV421 Mouse IgG1, k Isotype Control (Cat. No. 562438; Left Plot) or BD Horizon™ BV421 Mouse Anti-Human KLRG1 antibody (Cat. No. 568268/568269; Right Plot). After staining, the cells were treated with 1× BD Pharm Lyse™ lysing solution (Cat. No. 555899) to remove erythrocytes. Bivariate pseudocolor density plots showing the correlated expression of KLRG1 (or Isotype control) versus CD56 were derived from gated events based on side and forward light-scattering characteristics of intact lymphocyte populations. Flow cytometric analysis was performed using a BD BD Fortessa™ X-20 Flow Cytometer System and FlowJo™ software.
BD Horizon™ BV421 Mouse Anti-Human KLRG1
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Preparation And Storage
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Product Notices
- An isotype control should be used at the same concentration as the antibody of interest.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
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- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
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- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
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The Z7-205.rMAb monoclonal antibody specifically binds to KLRG1 (Killer cell Lectin-like Receptor G1), which is the homolog of the rat mast cell function-associated antigen (MAFA). KLRG1 is an inhibitory lectin-like type II transmembrane receptor containing a ITIM (Immunoreceptor Tyrosine-based Inhibitory Motif) cytoplasmic motif.. KLRG1 is expressed mainly as a homodimeric molecule consisting of two 30-38 kDa N-glycosylated subunits. Binding to its ligands E-, N- and R-cadherins prevents Akt phosphorylation and increases expression of cell cycle inhibitors. Human KLRG1 is expressed on a large subset of NK cells, lymphokine-activated killer (LAK) cells, adherent LAK (A-LAK) cells, subsets of activated CD8+ T lymphocytes, and small fractions of CD4+ and CD8+ T cells, but not mast cells (unlike the rat homolog, which is expressed on mast cells). KLRG1 expression is correlated with reduced proliferative capacity and effector functions of activated T lymphocytes and NK cells.
Development References (5)
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Akhmetzyanova I, Zelinskyy G, Littwitz-Salomon E, et al. CD137 Agonist Therapy Can Reprogram Regulatory T Cells into Cytotoxic CD4+ T Cells with Antitumor Activity.. J Immunol. 2016; 196(1):484-92. (Biology). View Reference
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Henson SM, Akbar AN. KLRG1--more than a marker for T cell senescence.. Age (Dordr). 2009; 31(4):285-91. (Biology). View Reference
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Levin MJ, Kroehl ME, Johnson MJ, et al. Th1 memory differentiates recombinant from live herpes zoster vaccines.. J Clin Invest. 2018; 128(10):4429-4440. (Biology). View Reference
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Wang D, Diao H, Getzler AJ, et al. The Transcription Factor Runx3 Establishes Chromatin Accessibility of cis-Regulatory Landscapes that Drive Memory Cytotoxic T Lymphocyte Formation.. Immunity. 2018; 48(4):659-674.e6. (Biology). View Reference
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Wu H, Tang X, Kim HJ, et al. Expression of KLRG1 and CD127 defines distinct CD8+ subsets that differentially impact patient outcome in follicular lymphoma.. J Immunother Cancer. 2021; 9(7):e002662. (Biology). View Reference
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