Anti nk1.1 pk136

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Anti-NK1.1 (PK136) is a laboratory reagent used for the identification and detection of NK1.1-positive cells in mice. It is a monoclonal antibody that specifically binds to the NK1.1 antigen expressed on natural killer cells and a subset of T cells in mice. This reagent can be used in various immunological techniques, such as flow cytometry, immunohistochemistry, and cell sorting, to study the role of NK1.1-positive cells in biological processes.

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NK1.1 (PK136) (18 citations) Anti-NK1.1 (17 citations) Anti-NK1.1 (clone PK136, (5 citations) PE-labeled anti-mouse NK1.1 (clone PK136) mouse IgG2aκ (2 citations) Anti-NK1.1 FITC (PK136) (2 citations)

4 protocols using anti nk1.1 pk136

Flow Cytometric Analysis of Murine NK Cells

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Spleen cells from BALB/c, C57BL/6, BALB.B6-CT-1, BALB.B6-CT-6, BALB.B6-CT-12 and BALB.B6-CT-13 mice were incubated with anti-CD16 antibody (Fc-block), washed and then stained with PE-conjugated anti-CD49b (DX-5) and PE-CY5-conjugated anti-TCR (H57-597) antibodies. Some samples were simultaneously stained with other antibodies such as FITC-conjugated anti-Ly49A (A1), Ly49D (4E5), Ly49G₂ (Cwy-e), Ly49I (YLI-90), anti-NKG2A/C/E (20d5) or anti-NK1.1 (PK136) for 1 h on ice (all antibodies and conjugates are from BD Pharmingen, San Diego, CA, USA). The cells were then washed twice, resuspended in PBS and analysed in a FACScalibur cytofluorometer (BD Biosciences, NJ) using CellQuest software. Viable lymphocytes were gated by forward and side scatter.

Hansen D.S., Ryg-Cornejo V., Ioannidis L.J., Chiu C.Y., Ly A., Nie C.Q., Scalzo A.A, & Schofield L. (2014). The Contribution of Natural Killer Complex Loci to the Development of Experimental Cerebral Malaria. PLoS ONE, 9(4), e93268.

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Flow Cytometric Analysis of Activated Immune Cells

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Cell surface staining was performed using the following antibodies: anti-CD11b (M1/70, Miltenyi Biotec), anti-CD11c (N418, BioLegend) anti-CD3 (17A2, eBioscience), anti-CD49b (DX5, eBioscience), anti-CD69 (H1.2 F3, eBioscience), anti-CD86 (GL-1, BioLegend), anti-MHC-II (M5/114.15.2, Miltenyi Biotec), anti-NK1.1 (PK136, BD Bioscience). Dead cells were excluded from analysis (positive for fixable viability dye, eBioscience). For detection of activated T cells splenocytes were stained with anti-CD4 (GK1.5, eBioscience), anti-CD8 (53–6.7, eBioscience), anti-CD43 (1B11, BioLegend) and anti-CD62L (MEL-14, eBioscience). Intracellular IFN-γ (XMG1.2, Miltenyi Biotec), IL-2 (JES6-5H4, Miltenyi Biotec) and TNF-α (MP6-XT22, BioLegend) staining was performed as described [28 (link),29 (link)]. Data were acquired on LSR II flow cytometer (BD Bioscience) and analyses were performed using FACSDiva (BD Bioscience) and Flow Jo (Tree Star, USA) software.

Gibbert K., Francois S., Sigmund A.M., Harper M.S., Barrett B.S., Kirchning C.J., Lu M., Santiago M.L, & Dittmer U. (2014). Friend retrovirus drives cytotoxic effectors through Toll-like receptor 3. Retrovirology, 11, 126.

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Multiparameter Flow Cytometry Analysis

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Equivalent cell numbers were treated with Fc block (BD Biosciences) and surface stained with combinations of anti-MHC II (M5/114.15.2), anti-CD11c (N418), anti-CD11b (M1/70), and anti-Ly6c (HK1.4) antibodies (eBiosciences) (neutrophils) or anti-Ly6g (1A8), anti-MHC class II (AF6–120.1), anti-CD11c (N418; eBioscience), anti-Ly6c (ER-MP20; AbD Serotech), anti-CD11b (M1/70; BD Biosciences), anti-CD4 (RM4.5; BD Biosciences), anti-NK1.1 (PK136; BD Biosciences), αβT cell receptor (H57–597; BD), anti-CD8 (53–6.7; eBioscience), anti-iNOS (M-19; Santa Cruz Biotechnology), or anti-TNF-α (MP6-XT22; BD Biosciences) (tipDCs, CD4⁺ T cells, CD8⁺ T cells) as previously described [37 (link),66 (link)]. CD8⁺ T cells were stained with tetramers specific for the D^bPB1-F2_62–70 epitope for 1 h at room temperature prior to surface staining with the monoclonal antibodies as previously described [66 (link)].

Meliopoulos V.A., Van de Velde L.A., Van de Velde N.C., Karlsson E.A., Neale G., Vogel P., Guy C., Sharma S., Duan S., Surman S.L., Jones B.G., Johnson M.D., Bosio C., Jolly L., Jenkins R.G., Hurwitz J.L., Rosch J.W., Sheppard D., Thomas P.G., Murray P.J, & Schultz-Cherry S. (2016). An Epithelial Integrin Regulates the Amplitude of Protective Lung Interferon Responses against Multiple Respiratory Pathogens. PLoS Pathogens, 12(8), e1005804.

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Identification of Immune Cell Populations

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The following Abs were purchased from BD Pharmingen: allophycocyanin (APC)-conjugated anti-CD49b (HM2), phycoerythrin (PE)-conjugated anti-Siglec-F (E50-2440), and anti-NK1.1 (PK136). Unlabeled anti-IL-3 (MP2-8F8) and rat IgG1 isotype (RTK2071); biotin-conjugated anti-CD49b (DX5), anti-CD123 (IL-3R, 5B11), anti-FcRI (MAR-1), anti-I-A/I-E (M5/114.15.2), American hamster IgG isotype (HTK888), and rat IgG2a isotype (RTK2758); PE-conjugated CD200R3 (Ba13), anti-IgE (RME-1), and anti-CD4 (RM4.5); APC-conjugated anti-Gr-1 (RB6-8C5), anti-B220 (RA3-6B2), and anti-F4/80 (BM8); PE/Cy7-conjugated anti-c-kit (2B8) and anti-CD45 (30-F11); Brilliant Violet 421-conjugated anti-CD3 (145-2C11) and anti-CD11c (N418); and APC/Cy7-conjugated streptavidin were from BioLegend. PE-conjugated CD11b Abs (M1/70) were obtained from eBioscience. Anti-STIM1 (D88E10), anti-STIM2 (4917), and anti--tubulin (2144) were from Cell Signaling Technology. TNP-specific IgE (IGEL-b4) and anti-CD16/32 (2.4G2) were prepared in our laboratory. TAM (T5648), DT (D0564), inactive mDT (D2189), EGTA, BAPTA-AM, and DPI were purchased from Sigma-Aldrich. Recombinant mouse IL-3 and IL-33 were obtained from BioLegend.

Yoshikawa S., Oh-Hora M., Hashimoto R., Nagao T., Peters L., Egawa M., Ohta T., Miyake K., Adachi T., Kawano Y., Yamanishi Y, & Karasuyama H. (2019). Pivotal role of STIM2, but not STIM1, in IL-4 production by IL-3-stimulated murine basophils. Science signaling, 12(576).

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