BV650 Hamster Anti-Mouse KLRG1

This product is the replacement for 740553.

BD Horizon™ BV650 Hamster Anti-Mouse KLRG1

Name: BV650 Hamster Anti-Mouse KLRG1
Brand: BD Horizon™
SKU: 569548

Clone 2F1

(RUO)

Multicolor flow cytometric analysis of KLRG1 expression on viable Mouse splenic leucocytes. C57BL/6 Mouse splenocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with PE Mouse Anti-Mouse NK-1.1 antibody (Cat. No. 553165/557391/561046) and with either BD Horizon™ BV650 Hamster IgG2, κ Isotype Control (Cat. No. 563860; Left Panel) or BD Horizon™ BV650 Hamster Anti-Mouse KLRG1 antibody (Cat. No. 569548; Right Panel) at 0.25 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) 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) leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

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Product Details

Brand: BD Horizon™

Alternative Name: Klrg1; Killer cell lectin-like receptor subfamily G member 1; MAFA; 2F1 Ag

Reactivity: Mouse (QC Testing)

Isotype: Syrian Hamster IgG2, κ

Immunogen: A-LAK from C57BL/6 mice

Application: Flow cytometry (Routinely Tested)

Concentration: 0.2 mg/ml

Entrez Gene ID: 50928

Storage Buffer: Aqueous buffered solution containing ≤0.09% sodium azide.

Regulatory Status: RUO

Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. 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 unconjugated antibody and free dye were removed.

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

Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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.
Since applications vary, each investigator should titrate the reagent to obtain optimal results.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
An isotype control should be used at the same concentration as the antibody of interest.
BD Horizon Brilliant Violet 650 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,455,613; 8,575,303; 8,354,239.
BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
Alexa Fluor™ is a trademark of Life Technologies Corporation.
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

Brilliant Stain Buffer RUO

Size 1000 Tests Cat No. 566349

Brilliant Stain Buffer Plus RUO

Size 1000 Tests Cat No. 566385

BV650 Hamster IgG2, κ Isotype Control RUO

Size 50 µg Cat No. 563860

Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO

Size 0.5 mg Cat No. 553142

View Details

569548 Rev. 1

Antibody Details

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.

569548 Rev. 1

Format Details

BV650

The BD Horizon Brilliant Violet™ 650 (BV650) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This tandem fluorochrome is comprised of a BV421 donor with an excitation maximum (Ex Max) of 406-nm and an acceptor dye with an emission maximum (Em Max) at 649-nm. BV650, driven by BD innovation, is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 650-nm (e.g., a 660/20-nm bandpass filter). The acceptor dye can be excited by the Red (628–640-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: BV650

Excitation Source: Violet 405 nm

Excitation Max: 406 nm

Emission Max: 649 nm

569548 Rev.1

Citations & References

Development References (3)

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
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
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

569548 Rev. 1

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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.