Cells were pre-incubated with human IgG (Baxter). We purchased the following specific mAbs from Miltenyi Biotec: CD3 (IgG2a), CD14 (IgG2a), CD56 (IgG1), CD314 (NKG2D) (IgG1) and CD107a (IgG1); from BD Biosciences IFN-γ (IgG1), CD73 (IgG1), Granzyme A (IgG1), CD44 (IgG2b) and RANK (IgG1); from Beckman Coulter-Immunotech CD337 (NKp30) (IgG1), CD335 (NKp46) (IgG1) and CD336 (NKp44) (IgG1); from R&D Systems ULBP1 (IgG2a), ULBP2 (IgG2a), ULPB3 (IgG2a) anti-IL8 (IgG1), IL15α (IgG2b), CD122 (IgG1) and CD132 (IgG2a); from Ancell CD29 (IgG1), CD105 (IgG1), CD106 (IgG1) and perforin (IgG2b); from Amgen ULBP-4 (IgG1); from BioLegend CD1a (IgG1), CD90 (IgG1) and RANKL (IgG2b); from Life Technologies Granzyme B (IgG1); from Santa Cruz biotechnology Vimentin (IgG1); from LifeSpan BioSciences Cytokeratin-7 (IgG1); from Abcam Desmin (IgG). CD146 (IgG2a), M7E22 mAb anti-CD54 (IgG1), D1–12 mAb anti-HLA-DR, W6.32 mAb anti-HLA-I (IgG2a), L95 mAb anti-PVR (IgG2a), L14 mAb anti-Nectin-2 (IgG2a) and BAM195 mAb anti-MICA (IgG1) were kindly provided by Pende D. (Genoa, Italy). We purchased secondary conjugated-specific mAbs from Invitrogen or Southern Biotech. All samples were analyzed on Gallios Flow Cytometer (IL-Beckman Coulter). Data analysis was done using FlowJo software.
Cd314 nkg2d
Manufactured by BD
The CD314 (NKG2D) is a receptor protein expressed on the surface of natural killer (NK) cells, CD8+ T cells, and other immune cells. It plays a role in the recognition and elimination of stressed or transformed cells. This product provides a tool for studying the function and expression of the CD314 receptor on various cell types.
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Lab products found in correlation
Rankl, by Thermo Fisher Scientific (1 mentions)
Desmin, by Abcam (1 mentions)
Ulbp1, by R&D Systems (1 mentions)
Cytokeratin 7, by Abcam (1 mentions)
Granzyme b, by Santa Cruz Biotechnology (1 mentions)
Vimentin, by LifeSpan BioSciences (1 mentions)
Ifn γ, by BD (1 mentions)
Facs canto 2 instrument, by BD (1 mentions)
Anti tim 3, by R&D Systems (1 mentions)
Cd159a nkg2a, by Beckman Coulter (1 mentions)
Cd158a, by BD (1 mentions)
Cd56 v450, by BD (1 mentions)
Cd122, by BD (1 mentions)
Diva software, by BD (1 mentions)
2 protocols using cd314 nkg2d
1
Multiparametric Flow Cytometry Profiling
2
Phenotyping of NK Cells by Flow Cytometry
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Fluorochrome‐labeled monoclonal antibodies specific for CD3 (V500) and CD56 (V450) (BD Biosciences, San Jose, CA) were used to identify NK (CD3negCD56pos) cells within the overall lymphocyte population. Anti‐NKR antibodies CD16, CD161, CD122, CD158a, CD158e, CD94, CD69, CD95 (FAS), CD226 (DNAM), CD253 (TRAIL), and CD314 (NKG2D) were purchased from BD Biosciences. Anti‐CD335 (NKp46), CD337 (NKp30), and CD159a (NKG2A) were purchased from Beckman Coulter (Indianapolis, IN). Anti‐CD158b and CD328 (Siglec‐7) were purchased from eBioscience (San Diego, CA). Anti‐Tim‐3 was purchased from R&D Systems (Minneapolis, MN).
Thawed PBMCs (1‐2 × 106) were stained for cell‐surface antigen expression at 4°C in the dark for 30 minutes. Samples were then washed twice in 2 mL phosphate‐buffered saline containing 1% bovine serum albumin and 0.01% sodium azide (Facs Wash; MilliporeSigma, St. Louis, MO) and subsequently fixed in 200 μL 1X BD stabilization fixative. Isotype‐matched control antibodies were used to determine background levels of staining. Multicolor flow cytometry was performed using a BD FACSCanto II instrument (BD Biosciences), compensated with single fluorochromes and analyzed using Diva software (BD Biosciences). Lymphocyte populations were identified by their characteristic forward scatter/side scatter properties.
Thawed PBMCs (1‐2 × 106) were stained for cell‐surface antigen expression at 4°C in the dark for 30 minutes. Samples were then washed twice in 2 mL phosphate‐buffered saline containing 1% bovine serum albumin and 0.01% sodium azide (Facs Wash; MilliporeSigma, St. Louis, MO) and subsequently fixed in 200 μL 1X BD stabilization fixative. Isotype‐matched control antibodies were used to determine background levels of staining. Multicolor flow cytometry was performed using a BD FACSCanto II instrument (BD Biosciences), compensated with single fluorochromes and analyzed using Diva software (BD Biosciences). Lymphocyte populations were identified by their characteristic forward scatter/side scatter properties.
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