Cd226

Manufactured by BioLegend

Sourced in United States

CD226, also known as DNAM-1, is a cell surface glycoprotein that functions as an activating receptor. It is expressed on the surface of various immune cells, including natural killer (NK) cells, T cells, and platelets.

Automatically generated - may contain errors

Lab products found in correlation

4 protocols using cd226

TIGIT Expression on T Cell Subsets in Healthy Donors

Check if the same lab product or an alternative is used in the 5 most similar protocols

Example 12

TIGIT expression on T cell subsets in healthy donors was evaluated by flow cytometery. Frozen PBMC from 12 healthy donors were purchased from Astarte Biologics (Bothell, Wash.) and Folio Conversant (Huntsville, Ala.). Donor ages ranged from 21 to 74 with a median age of 48. Cells were stained with a viability dye and the following anti-human antibodies following Fc receptor blocking: CD3, CD96, CD226, CCR7, CD25, CD127, CD45RA, CD8, CD4, CD226, CD155 (Biolegend), and TIGIT (eBioscience). Stained cells were then analyzed on an Attune Nxt flow cytometer (Life Technologies). Treg subsets were defined by appropriate expression of CD4, CD25 and CD127) (CD4⁺CD25⁺CD127^lo). CD4⁺ and CD8⁺ effector memory (T_EM; CD45RA⁻CCR7⁻), CD45RA⁺ effector memory (T_EMRA; CD45RA⁺CCR7⁻), central memory (T_CM; CD45RA⁻CCR7⁺), and naïve (T_N; CD45RA⁺CCR7⁺) cell subsets were defined according to CD45RA and CCR7 expression. As shown in FIG. 3, TIGIT expression is highest on Tregs, while the activating receptor CD226 expression is lower on both memory and naïve CD8 T cell subsets and shows similar levels to naïve CD4 T cell subsets. CD155 expression is mostly absent among T cells but was seen on antigen presenting cells.

US11401339B2. Anti-TIGIT antibodies (2022-08-02). Seagen, Inc. [US]. Inventors: Julia C. Piasecki [US], Courtney Beers [US], Scott Peterson [US], Bianka Prinz [US], Shyra Gardai [US].

+ Open protocol

+ Expand

Comprehensive Immune Cell Profiling by FACS

Check if the same lab product or an alternative is used in the 5 most similar protocols

Fluorescence-activated cell sorting studies were performed using: FITC-, PE-, PE-Cy5, APC-Cy7, PE-Cy7, Pacific blue-, or APC-conjugated anti-human antibodies to: CD3 (clone#: HIT3a, 1:50), CD8 (SK1 or RPA-T8, 1:20), IL-17 (BL168, 1:20), IFNγ (4 S.B3, 1:10), IL-10 (JES3-19F1, 1:10), IL-4 (8D4-8, 1:10), FOXP3 (206D, 1:10), CD39 (A1, 1:20), CD73 (AD2, 1:20), CD28 (CD28.2, 1:20), CD45RA (HI100, 1:20), CD226 (TX25, 1:20), CXCR3 (G025H7, 1:20), Tim3 (F38-2E2, 1:20), CCR6 (G034E3, 1:20), Perforin (B-D48, 1:20), Granzyme B (GB11, 1:20) and CCR7 (g043h7, 1:20) from BioLegend (San Diego, CA, USA); CD39 (BU61, 5 μg ml⁻¹) from Ancell Corporation (Bayport, MN, USA); Annexin V from BD Biosciences (1:20; Franklin Lakes, NJ, USA); phospho-JNK (Thr183/Tyr185; G9, 1:20) and phospho-NFκB p65 (Ser536; 93H1, 1:20) from Cell Signalling Technology (Danvers, MA, USA); and Isotype control antibodies from Ancell Corporation or eBioscience (San Diego, CA, USA).
Antibodies used for western blot included the following: phospho-PI3K p85 (Tyr458; #4228, 1:800), phospho-Akt (Ser473; #9271, 1:1,000), phospho-mTOR (Ser2448; #2971, 1:1,000), mTOR (#2972, 1:1,000), pJNK (Thr183/Tyr185; #9251, 1:1,000) and phospho-NFκB p65 (Ser536; #3031, 1:1,000), from Cell Signaling Technology (Danvers, MA, USA); NOX2 (ab31092, 1 μg ml⁻¹) and β-actin (AC-15, #ab6276, 1:40,000) from Abcam (Cambridge, MA, USA).

Bai A., Moss A., Rothweiler S., Serena Longhi M., Wu Y., Junger W.G, & Robson S.C. (2015). NADH oxidase-dependent CD39 expression by CD8+ T cells modulates interferon gamma responses via generation of adenosine. Nature Communications, 6, 8819.

+ Open protocol

+ Expand

Comprehensive Immune Profiling of T Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

After T cell activation, T cells were stained with directly labeled antibodies (or isotype controls) as follows: CD46 (FITC, clone: MEM-258, Biolegend), CD11a (PE, clone HI111, Biolegend), CD69 (FITC, clone FN50, Biolegend), CD99 (PE, clone 3B2/TA8, Biolegend), CD29 (PE, clone: TS2/16, Biolegend), CD49d (APC, clone: 9F10, Biolegend), CXCR3 (PE human, Miltenyi Biotech), CD226 (APC, clone 11A8, Biolegend), CD146 (APC, clone P1H12, Biolegend), CD166 (PE, clone 3A6, Biolegend), TIGIT (APC, clone: A15 15 36, Biolegend). Cells were analyzed using a FACSCalibur or a 5 laser LST Fortessa (BD Biosciences). A live-dead marker (LIVE/DEAD™ Fixable Far Red Dead Cell Stain, Thermofisher) was added to the cells to exclude dead cells. The delta geometric mean is shown (MFI staining – MFI isotype). Representative staining are shown in Supplementary Figure S1. For the cohort of patients supplemented with vitamin D, PBMCs were analyzed with a panel of antibodies consisting of CD3-AF700, CD4-FITC, CD25-BV605, CD127-BV421, CD45RA-BV510, Foxp3-PE, CD46-PE/cy7, CD226-PE-dazzle 594, CD146-APC, CD162-BV711, CD6-BV785. Viable cells were selected using the NIR-ZOMBIE APC-Cy7 (Biolegend) (Supplementary Figure S1C). Cells were analyzed using a Fortessa (BD Biosciences) and analysis was done using FlowJo and t-SNE clustering.

Killick J., Hay J., Morandi E., Vermeren S., Kari S., Angles T., Williams A., Damoiseaux J, & Astier A.L. (2020). Vitamin D/CD46 Crosstalk in Human T Cells in Multiple Sclerosis. Frontiers in Immunology, 11, 598727.

+ Open protocol

+ Expand

Multiparameter Analysis of Immune Checkpoint Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

Alexa Fluor® 647 antihuman TIGIT mAb (clone MBSA43) was obtained from BioLegend® (San Diego, CA, USA). The antibodies to evaluate CD155 (Alexa Fluor® 647, clone SKII.4), CD112 (PE, clone R2.525), PDL-1 (PerCP-Cy5.5, clone 29E.2A3), NKp30 (PE, clone P30-15), NKp44 (PE, clone P44-8), NKp46 (PE, clone 9E2), CD226 (PE, clone DX11), and NKG2D (PE, clone 1D11) were obtained from BioLegend® and MICA (PE, clone 159227), MICB (FITC, clone 236511), ULBP1 (PerCP, clone 170818), ULBP2-5-6 (PE, clone 165903), and B7-H6 (APC, clone 875002) from R&D Systems (Minneapolis, MN, USA). In addition, Ultra-LEAF ™ purified antihuman TIGIT antibody (clone A15153A, mouse IgG2a, BioLegend®) and anti-IL6R antibody (Tocilizumab/Actemra®; Roche, Basil, Switzerland) were used for functional blocking assays at 50 μg/mL and 10 μg/mL, respectively. Stattic (STAT-3 inhibitor; CAS 19983-44-9) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). This reagent was reconstituted in dimethyl sulfoxide (DMSO, Sigma-Merck, Darmstadt, Germany) as 50 mM stock solutions and stored at −20 °C until use.

González-Ochoa S., Tellez-Bañuelos M.C., Méndez-Clemente A.S., Bravo-Cuellar A., Hernández Flores G., Palafox-Mariscal L.A., Haramati J., Pedraza-Brindis E.J., Sánchez-Reyes K, & Ortiz-Lazareno P.C. (2022). Combination Blockade of the IL6R/STAT-3 Axis with TIGIT and Its Impact on the Functional Activity of NK Cells against Prostate Cancer Cells. Journal of Immunology Research, 2022, 1810804.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!