Multiparameter flow cytometric analysis of KLRG1 expression on Human peripheral blood leukocyte populations. Human whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leukocytes were washed, preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219) and stained with Alexa Fluor™ 647 Mouse Anti-Human NCAM-1 (CD56) antibody (Cat. No. 563443; Top Plots) and with either BD Horizon™ RB613 Mouse IgG1, κ Isotype Control (Cat. No. 571106; Left Plots) or BD Horizon™ RB613 Mouse Anti-Human KLRG1 antibody (Cat. No. 571358/571360; Right Plots). Top Plots - The bivariate pseudocolor density plot showing the correlated expression of KLRG1 (or Ig Isotype control staining) versus NCAM-1 (CD56) was derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Bottom Plots - The bivariate pseudocolor density plot showing KLRG1 expression (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of viable leukocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software.
Multiparameter flow cytometric analysis of KLRG1 expression on Human peripheral blood leukocyte populations. Human whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leukocytes were washed, preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219) and stained with Alexa Fluor™ 647 Mouse Anti-Human NCAM-1 (CD56) antibody (Cat. No. 563443; Top Plots) and with either BD Horizon™ RB613 Mouse IgG1, κ Isotype Control (Cat. No. 571106; Left Plots) or BD Horizon™ RB613 Mouse Anti-Human KLRG1 antibody (Cat. No. 571358/571360; Right Plots). Top Plots - The bivariate pseudocolor density plot showing the correlated expression of KLRG1 (or Ig Isotype control staining) versus NCAM-1 (CD56) was derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Bottom Plots - The bivariate pseudocolor density plot showing KLRG1 expression (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of viable leukocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software.
Product Details
BD Horizon™
Killer cell lectin-like receptor subfamily G member 1; MAFA
Human (QC Testing)
Mouse IgG1, κ
Hepa 1-6 cells transfected with hKLRG1 DNA
Flow cytometry (Routinely Tested)
5 µl/test
10219
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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.
Product Notices
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
- 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.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
- CF™ is a trademark of Biotium, Inc.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Human BD Fc Block™ RUO
Size
50 µg
Cat No.
564219
RB613 Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
571106
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
Lysing Solution 10X Concentrate IVD
Size
100 mL
Cat No.
349202
571358 Rev. 1
Antibody Details
Z7-205.rMAb
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.
571358 Rev. 1
Format Details
RB613
The BD Horizon RealBlue™ 613 (RB613) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 492-nm and an emission maximum (Em Max) at 613-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB613 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with reduced excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB613 can be used as an alternative to PE-CF594 and we recommend using an optical filter centered near 610-nm (eg, a 610/20-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PE-CF594. RB613 is on average brighter than PE-CF594 off the blue laser.
RB613
Blue 488 nm
492 nm
613 nm
571358 Rev.1
Citations & References
View product citations for antibody "571358" on CiteAb
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
571358 Rev. 1
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.