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RY610 Hamster Anti-Mouse KLRG1
RY610 Hamster Anti-Mouse KLRG1
Multicolor flow cytometric analysis of KLRG1 expression on viable Mouse splenic leukocytes. C57BL/6 Mouse splenocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553142]. The cells were then stained with BD Horizon™ BV421 Mouse Anti-Mouse NK-1.1 antibody (Cat. No. 562921) and with either BD Horizon™ RY610 Hamster IgG1, κ Isotype Control (Cat. No. 571251 was used as a representative Hamster IgG Isotype Control; Left Plot) or BD Horizon™ RY610 Hamster Anti-Mouse KLRG1 antibody (Cat. No. 571238/571302; Right Plot) at 0.5 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of KLRG1 (or Ig Isotype control staining) versus NK-1.1 was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) leukocytes. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
Multicolor flow cytometric analysis of KLRG1 expression on viable Mouse splenic leukocytes. C57BL/6 Mouse splenocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553142]. The cells were then stained with BD Horizon™ BV421 Mouse Anti-Mouse NK-1.1 antibody (Cat. No. 562921) and with either BD Horizon™ RY610 Hamster IgG1, κ Isotype Control (Cat. No. 571251 was used as a representative Hamster IgG Isotype Control; Left Plot) or BD Horizon™ RY610 Hamster Anti-Mouse KLRG1 antibody (Cat. No. 571238/571302; Right Plot) at 0.5 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of KLRG1 (or Ig Isotype control staining) versus NK-1.1 was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) leukocytes. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
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BD Horizon™
Klrg1; Killer cell lectin-like receptor subfamily G member 1; MAFA; 2F1 Ag
Mouse (QC Testing)
Syrian Hamster IgG2, κ
A-LAK from C57BL/6 mice
Flow cytometry (Routinely Tested)
0.2 mg/ml
50928
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

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. 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.
  5. 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.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  8. CF™ is a trademark of Biotium, Inc.
  9. For U.S. patents that may apply, see bd.com/patents.
571302 Rev. 1
Antibody Details
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2F1

The 2F1 monoclonal antibody specifically binds to KLRG1 (Killer cell Lectin-like Receptor G1), which is the mouse homolog of the rat mast cell function-associated antigen (MAFA), on all mouse strains tested (eg, AKR/J, BALB/c, C3H/HeN, C3H.SW, C57BL/6, DBA/1, SJL, 129/J). Unlike rat MAFA, which is expressed on mast cells, mouse 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. The expression of KLRG1 is correlated with reduced proliferative capacity of activated T lymphocytes or reduced effector functions of activated NK cells. KLRG1 plays a role in the regulation of leucocytes of both the innate and adaptive immune system. The 2F1 mAb reportedly stains the rat basophilic leukemia cell line, RBL-2H3, which is known to express MAFA. The KLRG1 protein is an inhibitory lectin-like type II transmembrane receptor containing a cytoplasmic motif similar to ITIM (Immunoreceptor Tyrosine-based Inhibitory Motif); its ligand has not been identified KLRG1 is expressed mainly as a homodimeric molecule consisting of two N-glycosylated subunits of approximately 30-38 kDa. The level of KLRG1 expression is reduced in MHC class I-deficient mice, although direct binding of KLRG1 to MHC class I antigens could not be detected. Crosslinking of KLRG1 by 2F1 mAb reduces TCR-mediated Ca++ mobilization and cytotoxic responses (but not IFN-γ production) by CD8+ T cells and inhibits IFN-γ and TNF production and redirected lysis by NK cells.

571302 Rev. 1
Format Details
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RY610
The BD Horizon RealYellow™ 610 (RY610) Dye is part of the BD® family of yellow-green dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 557-nm and an emission maximum (Em Max) at 610-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RY610 can be used on both spectral and conventional cytometers and is designed to be excited by the Yellow-Green laser (561-nm) with minimal excitation by the 488-nm blue laser. For conventional instruments equipped with a Yellow-Green laser (561-nm), RY610 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 Yellow-Green laser (561-nm), it can be used in conjunction with PE-CF594.
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RY610
Yellow-Green 561 nm
557 nm
610 nm
571302 Rev.1
Citations & References
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View product citations for antibody "571302" on CiteAb

Development References (3)

  1. Beyersdorf NB, Ding X, Karp K, Hanke T. Expression of inhibitory "killer cell lectin-like receptor G1" identifies unique subpopulations of effector and memory CD8 T cells. Eur J Immunol. 2001; 31(12):3443-3452. (Clone-specific: Bioassay, Blocking, Functional assay). View Reference
  2. Corral L, Hanke T, Vance RE, Cado D, Raulet DH. NK cell expression of the killer cell lectin-like receptor G1 (KLRG1), the mouse homolog of MAFA, is modulated by MHC class I molecules. Eur J Immunol. 2000; 30(3):920-930. (Immunogen: Flow cytometry, Immunoprecipitation). View Reference
  3. Hanke T, Corral L, Vance RE, Raulet DH. 2F1 antigen, the mouse homolog of the rat "mast cell function-associated antigen", is a lectin-like type II transmembrane receptor expressed by natural killer cells. Eur J Immunol. 1998; 28(12):4409-4417. (Clone-specific: Flow cytometry). View Reference
571302 Rev. 1

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