PerCP-Cy™5.5 Hamster Anti-Mouse KLRG1
Clone 2F1 (RUO)
- Brand BD Pharmingen™
- Alternative Name Klrg1; Killer cell lectin-like receptor subfamily G member 1; MAFA; 2F1 Ag
- Concentration 0.2 mg/ml
- Isotype Syrian Hamster IgG2, κ
- Reactivity Mouse (QC Testing)
Flow cytometry (Routinely Tested)
- Immunogen A-LAK from C57BL/6 mice
- Entrez Gene ID 50928
- Storage Buffer Aqueous buffered solution containing ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
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.
PerCP-Cy™5.5 is a tandem conjugate that combines PerCP with a cyanine dye. PerCP-Cy5.5 is not subject to photobeaching like PerCP and can be used with stream-in-air flow cytometers. It has less Fc receptor–mediated nonspecific staining than PE-Cy5. Additionally, the PerCP-Cy5.5 tandem conjugate is not as susceptible to fixative or light instability compared to APC-Cy7 and PE-Cy7.
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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 with PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Cy is a trademark of Amersham Biosciences Limited. This conjugated product is sold under license to the following patents: US Patent Nos. 5,486,616; 5,569,587; 5,569,766; 5,627,027.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- 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.
- PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
- PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- This product is subject to proprietary rights of Amersham Biosciences Corp. and Carnegie Mellon University and made and sold under license from Amersham Biosciences Corp. This product is licensed for sale only for research. It is not licensed for any other use. If you require a commercial license to use this product and do not have one return this material, unopened to BD Biosciences, 10975 Torreyana Rd, San Diego, CA 92121 and any money paid for the material will be refunded.
- Although hamster immunoglobulin isotypes have not been well defined, BD Biosciences Pharmingen has grouped Armenian and Syrian hamster IgG monoclonal antibodies according to their reactivity with a panel of mouse anti-hamster IgG mAbs. A table of the hamster IgG groups, Reactivity of Mouse Anti-Hamster Ig mAbs, may be viewed at http://www.bdbiosciences.com/documents/hamster_chart_11x17.pdf.