Skip to main content Skip to navigation
PerCP-Cy™5.5 Mouse Anti-Mouse NK-1.1
PerCP-Cy™5.5 Mouse Anti-Mouse NK-1.1

Two-color analysis of NK-1.1 expression on lymphokine-activated killer (LAK) cells. LAK cells from C57BL/6 mice were stained with FITC-conjugated anti-mouse CD244.2 mAb 2B4 (Cat. No. 553305) and either PerCP-Cy5.5-conjugated mouse IgG2a κ isotype control mAb G155-178 (Cat. No. 551115, left panel) or PerCP-Cy5.5-conjugated mAb PK136 (right panel). Viable blasts were gated according to their light-scatter profile. Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.

Two-color analysis of NK-1.1 expression on lymphokine-activated killer (LAK) cells. LAK cells from C57BL/6 mice were stained with FITC-conjugated anti-mouse CD244.2 mAb 2B4 (Cat. No. 553305) and either PerCP-Cy5.5-conjugated mouse IgG2a κ isotype control mAb G155-178 (Cat. No. 551115, left panel) or PerCP-Cy5.5-conjugated mAb PK136 (right panel). Viable blasts were gated according to their light-scatter profile. Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.

Product Details
Down Arrow Up Arrow


BD Pharmingen™
Klrb1b, CD161b, Nkrp1b; Klrb1c, CD161c, NK1.1, Nkrp1c
Mouse (QC Testing)
Mouse C3H x BALB/c IgG2a, κ
Mouse NK-1+ Spleen and Bone Marrow Cells
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_394052
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 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. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

Recommended Assay Procedures

PerCP has been reported to undergo significant photobleaching, the magnitudeof which increases as laser power is increased or beam focus is narrowed. For tandem conjugates incorporating PerCP (e.g., PerCP-Cy5.5), the excitation and emission properties of PerCP and the kinetics of energy exchange between the fluorochromes of the tandem dye may limit their effectiveness on high speed and/or sorting flow cytometers. Therefore, for third color flow cytometric analysis using ≥ 25-mW laser power, we recommend the PE-Cy7-conjugated reagent (Cat. No. 552878).

PerCP-Cy5.5 is a tandem fluorochrome composed of peridinin chlorophyll protein (PerCP), which is excited by the 488-nm line of an Argon ion laser and serves as the energy donor, coupled to the cyanine dye Cy5.5™, which acts as the energy acceptor and fluoresces at 695nm.

It is recommended that a 712/20-nm band-pass filter be used with stream-in-air instruments such as the BD FACStar™ and BD FACSVantage™ flow cytometry systems.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. 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.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. 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.
  7. 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.
  8. 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.
561111 Rev. 1
Antibody Details
Down Arrow Up Arrow
PK136

In the mouse, at least three members of the Klrb (Killer cell lectin-like receptor, subfamily b; formerly NKR-P1) gene family have been identified (Klrb1a/NKR-P1A, Klrb1b/NKR-P1B, and Klrb1c/NKR-P1C); but in the human gene family, a single homologue has been designated KLRB1, NKR-P1A, or CD161. The KLRB1/NKR-P1 family of proteins are type-II-transmembrane C-type lectin receptors. KLRB1C/NKR-P1C activates NK-cell cytotoxicity, while KLRB1B/NKR-P1B functions as an inhibitory receptor. KLRB1B/NKR-P1B protein has intracellular Immunoreceptor Tyrosine-based Inhibitory Motif (ITIM), while KLRB1C/NKR-P1C lacks ITIM and activates via association with Fc Receptor γ chain. Strikingly, KLRB1B/NKR-P1B and KLRB1C/NKR-P1C share 96% amino acid sequence identity in their extracellular C-type lectin domains. The PK136 antibody reacts with the NK-1.1 surface antigen (CD161c) encoded by the Klrb1c/NKR-P1C gene expressed on natural killer (NK) cells in selected strains of mice (eg, C57BL, FVB/N, NZB, but not A, AKR, BALB/c, CBA/J, C3H, C57BR, C58, DBA/1, DBA/2, NOD, SJL, 129) and the CD161b antigen encoded by the Klrb1b/NKR-P1B gene expressed only on Swiss NIH and SJL mice, but not on C57BL/6. Expression of KLRB1C/NKR-P1C protein is correlated with the ability to lyse tumor cells in vitro and to mediate rejection of bone marrow allografts. The NK-1.1 marker is useful in defining NK cells; however, the antigen is also expressed on a rare, specialized population of T lymphocytes (NK-T cells) and some cultured monocytes. Plate-bound PK136 mAb, in combination with low concentrations of IL-2, induces proliferation of a subset of NK cells.

561111 Rev. 1
Format Details
Down Arrow Up Arrow
PerCP-Cy5.5
PerCP-Cy5.5 dye is part of the BD blue family of dyes. This tandem fluorochrome is comprised of a fluorescent protein complex (PerCP) with an excitation maximum (Ex Max) of 482 nm and an acceptor dye with an emission maximum (Em Max) at 676 nm. PerCP-Cy5 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 680 nm (e.g., a 695/40 nm bandpass filter). The donor dye can be partially excited by the Violet (405-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.
altImg
PerCP-Cy5.5
Blue 488 nm
482 nm
676 nm
561111 Rev.1
Citations & References
Down Arrow Up Arrow

Development References (5)

  1. Carlyle JR, Martin A, Mehra A, Attisano L, Tsui FW, Zuniga-Pflucker JC. Mouse NKR-P1B, a novel NK1.1 antigen with inhibitory function. J Immunol. 1999; 162(10):5917-5923. (Clone-specific: Immunoprecipitation). View Reference
  2. Greimers R, Trebak M, Moutschen M, Jacobs N, Boniver J. Improved four-color flow cytometry method using fluo-3 and triple immunofluorescence for analysis of intracellular calcium ion ([Ca2+]i) fluxes among mouse lymph node B- and T-lymphocyte subsets. Cytometry. 1996; 23(3):205-217. (Biology). View Reference
  3. Koo GC, Peppard JR. Establishment of monoclonal anti-Nk-1.1 antibody. Hybridoma. 1984; 3(3):301-303. (Immunogen: Cytotoxicity, Flow cytometry). View Reference
  4. Kung SK, Su RC, Shannon J, Miller RG. The NKR-P1B gene product is an inhibitory receptor on SJL/J NK cells. J Immunol. 1999; 162(10):5876-5887. (Clone-specific: Blocking). View Reference
  5. Reichlin A, Yokoyama WM. Natural killer cell proliferation induced by anti-NK1.1 and IL-2. Immunol Cell Biol. 1998; 76(2):143-152. (Clone-specific: Induction). View Reference
View All (5) View Less
561111 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.