Nkg2a c e

Manufactured by BD

The NKG2A/C/E is a lab equipment product that detects the presence and expression levels of the NKG2A, NKG2C, and NKG2E receptors on cells. It provides a tool for researchers to study the role of these receptors in immune system function and response.

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3 protocols using nkg2a c e

Comprehensive Multicolor Flow Cytometry Protocol

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Single-cell suspensions were stained with the appropriate monoclonal antibody in PBS containing 2% FCS. When necessary, intracellular staining was performed by use of the FoxP3/Transcription Factor Staining Buffer Set (eBioscience) according to the manufacturer's instructions. Fortessa, FACS Verse, and ARIAIII (BD Biosciences) were used for cell sorting and analysis. Antibodies specific for NK1.1 (PK136; 1:100), CD19 (1D3; 1:400), CD3 (17A2; 1:400 or REA641; Miltenyi Biotec; 1:150); CD122 (TM-β1; 1:200), NKp46 (29A1.4; 1:100), KLRG1 (2F1; 1:200), CD27 (SB/199; 1:200), CD11b (M1/70; 1:200), IL-7R (A7R34; eBioscience; 1:200) CD49b (DX5; 1:100), CD49a (Ha31/8; 1:200) Ly49H (3D10; 1:200) Ly49D (4E5; 1:200), NKG2D (C4; 1:200), NKG2A/C/E (20d5; 1:200), Ly49C/I (5e6; 1:100), CD107a (104B; 1:100), and IFN-γ (XMG1.2; 1:100), DNAM-1 (10E5; 1:200); Ki-67 (AF488; 1:50) were from BD Pharmingen unless stated otherwise.

Delconte R.B., Guittard G., Goh W., Hediyeh-Zadeh S., Hennessy R.J., Rautela J., Davis M.J., Souza-Fonseca-Guimaraes F., Nunès J.A, & Huntington N.D. (2020). NK Cell Priming From Endogenous Homeostatic Signals Is Modulated by CIS. Frontiers in Immunology, 11, 75.

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Comprehensive Isolation and Characterization of Diverse Immune Cell Populations

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Single-cell suspensions were prepared from liver, spleen, bone marrow, uterus, salivary gland, thymus, and lung tissues. Liver lymphocytes were isolated at the interphase of a 40/60% Percoll gradient (1 (link)). Uterus and salivary gland were digested in the presence of DNase I and collagenase D or liberase TL (5 (link), 7 (link)), while lung digestion additionally incorporated trypsin inhibitor. Cells were stained with a LIVE/DEAD fixable dead cell stain kit (Invitrogen) and blocked with antibodies against CD16/CD32 (BD) prior to staining with antibodies for CD3, CD11b, CD27, CD43, CD122, human CD5, integrin α_v, KLRG1, Ly49G2, Ly49H, NKp46, and TRAIL (eBioscience); CD19, CD49a, CD49b, CD127, Ly49D, Ly-6G/Ly-6C (Gr-1), NK1.1, NKG2A/C/E, and TER-119 (BD). Cells were treated with cytofix/cytoperm kits prior to staining with antibodies for the transcription factors T-bet (eBioscience) and Eomes (eBioscience) or the cytokine TNF-α (BD).

Pikovskaya O., Chaix J., Rothman N.J., Collins A., Chen Y.H., Scipioni A.M., Vivier E, & Reiner S.L. (2016). Eomesodermin is Sufficient to Direct Type 1 Innate Lymphocyte Development into the Conventional Natural Killer Lineage. Journal of immunology (Baltimore, Md. : 1950), 196(4), 1449-1454.

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Isolation and Characterization of Murine Cortical Cells

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Cortices were collected from P5 to P8 pups in DMEM/F12 buffered with Hepes (15mM), and subcortical structures and meninges were removed. The cortex from one animal was chopped using sharp forceps and digested in 1 mL papain (4U/mL) and DNase (50 U/mL) in Hank’s balanced salt solution (HBSS, with cations) at 37 °C for 15 min. Each sample was triturated 10 times, filtered through a 70-µm filter, and washed with warm culture media (DMEM/F12 with 10% FBS and Anti-Anti). Pellets were resuspended in 15 mL of culture media and plated in T75 tissue-culture-treated flasks. A full media change was given ∼24 h after plating and half-medium changes (“feedings”) were given after 1 wk and every third subsequent day.
Cells were collected after 2 wk by gentle scraping, washed, and prepared for FACS analysis. Staining was performed with an antibody mixture that included antibodies (1:1,000) against the following: CD45, CD11b, Glast, NKG2A/C/E (all from BD Biosciences), and Zombie-Aqua viability dye (BioLegend). The cells were analyzed on a Bio-Rad ZE5 Cell Analyzer.

Marin I.A., Gutman-Wei A.Y., Chew K.S., Raissi A.J., Djurisic M, & Shatz C.J. (2022). The nonclassical MHC class I Qa-1 expressed in layer 6 neurons regulates activity-dependent plasticity via microglial CD94/NKG2 in the cortex. Proceedings of the National Academy of Sciences of the United States of America, 119(23), e2203965119.

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