Anti cd69 apc cy7

Manufactured by BD

The Anti-CD69 APC-Cy7 is a fluorescent-conjugated antibody that binds to the CD69 surface antigen. CD69 is a type II transmembrane protein that is rapidly induced on the surface of activated lymphocytes, including T cells, B cells, and natural killer cells. The APC-Cy7 fluorochrome is used to label the anti-CD69 antibody, providing a specific signal for flow cytometric detection and analysis.

Automatically generated - may contain errors

Lab products found in correlation

5 protocols using anti cd69 apc cy7

Cytotoxicity and Flow Cytometry Analysis

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

Analysis was done using a BD fluorescence-activated cell sorting (FACS) Canto II Flow Cytometer and BD LSRII Flow Cytometer (BD Biosciences, San Jose, CA). Data was analyzed using the BD FACSDiva software. Antibodies used included anti-CD69 APC-Cy7, anti-CD5 PerCP/Cy5.5, antihuman CD45 PE (BD Biosciences) and anti-myc tag fluorescein isothiocyanate (FITC,Abcam, Cambridge, MA). For the cytotoxicity studies, target cells were stained with the membrane dye PKH26 and cell death was assessed using 7-AAD (described below). Flow sorting for GFP was performed using a BD FACS Aria II Cell Sorter (BD Biosciences).

Moot R., Raikar S.S., Fleischer L., Querrey M., Tylawsky D.E., Nakahara H., Doering C.B, & Spencer H.T. (2016). Genetic engineering of chimeric antigen receptors using lamprey derived variable lymphocyte receptors. Molecular Therapy Oncolytics, 3, 16026-.

+ Open protocol

+ Expand

Multicolor Flow Cytometry Analysis

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

Analysis was performed using a BD LSRII Flow Cytometer (BD Biosciences, San Jose, CA). Data were analyzed using FCS Express 6 software. Antibodies used included anti-CD5 PerCP/Cy5.5, anti-CD3 BV421, anti-γδ T cell receptor (TCR) phycoerythrin (PE) and anti-CD69 APC-Cy7 (BD Biosciences, San Jose, CA). CD5-Fc fusion protein (G&P Biosciences, Santa Clara, CA) and CD19-Fc fusion protein (ACROBiosystems, Newark, DE) were used to detect anti-CD5 constructs and anti-CD19 constructs, respectively, with a secondary anti-IgG Fc antibody (Jackson Immunoresearch Laboratories, West Grove, PA), as previously described.⁶ (link) Violet Proliferation Dye 450 (VPD450) was used to label the target cells in the cytotoxicity and co-culture studies, and cell death was assessed using eFluor 780 (described below). Degranulation of γδ T cells was detected using anti-CD107a APC (BD Biosciences, San Jose, CA).

Fleischer L.C., Becker S.A., Ryan R.E., Fedanov A., Doering C.B, & Spencer H.T. (2020). Non-signaling Chimeric Antigen Receptors Enhance Antigen-Directed Killing by γδ T Cells in Contrast to αβ T Cells. Molecular Therapy Oncolytics, 18, 149-160.

+ Open protocol

+ Expand

Membrane Phenotype of Electroporated DCs

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

DC were membrane-stained with monoclonal anti-IL-15-PE antibody (R&D, Ref: LIT0713051) 2h, 4h, 8h, 24h, 48h and 72h after electroporation of the DC and analyzed on a FACScan flow cytometer (BD). In other experiments, purified NK cells were cocultured at a 5:1 ratio with autologous DC in IMDM + 10% FBS directly after DC electroporation and membrane-stained with anti-CD11c-V450 (BD; Ref: 560369), anti-NKp30-AF360 (BD; Ref: 558408), anti-NKp44-PE (BD; Ref: 558563), anti-NKp46-APC (BD; Ref: 557940), anti-NKG2D-PE (BD; Ref: 557940), anti-CD56-FITC (BD; Ref:345811) and anti-CD69-APC-Cy7 (BD; Ref: 557756) mABs 48h after initiation of cocultures. 7-aminoactinomycin D (7-AAD; BD; Ref: 51-68981E) was used to distinguish between viable and dead cells. Samples were measured on a FACSAria II flow cytometer (BD).

Van den Bergh J., Willemen Y., Lion E., Van Acker H., De Reu H., Anguille S., Goossens H., Berneman Z., Van Tendeloo V, & Smits E. (2015). Transpresentation of interleukin-15 by IL-15/IL-15Rα mRNA-engineered human dendritic cells boosts antitumoral natural killer cell activity. Oncotarget, 6(42), 44123-44133.

+ Open protocol

+ Expand

Comprehensive Immunophenotyping of NK Cells

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

Thawed PBMC were stained with ethidium monoazide (EMA), anti-CD19-PeCy5, anti-CD3-AlexaFluor700 (both from BD Biosciences), and with either anti-CD14-V711 (Biolegend) or anti-CD14-PeCy5 (AbD Serotec, Raleigh, NC) to exclude dead cells, T cells, B cells and monocytes. NK cells were identified using anti-CD56-PeCy7 (BD Biosciences). FITC-conjugated antibodies against CD122, CXCR3 (R&D Systems), CD69 and HLA-DR (BD Biosciences), PE-conjugated antibodies against TRAIL, CD300 (BD Biosciences), NKG2A, NKG2D, and NKp44 (Beckman Coulter, Brea, CA), and APC-conjugated antibodies against CD85j (eBiosciences, San Diego, CA), CCR5 (BD Biosciences), NKG2C (R&D Systems), NKp30 and NKp46 (Miltenyi Biotec, Auburn, CA) were added. Liver-infiltrating lymphocytes were stained with anti-TRAIL-PE, anti-CD69-APC/Cy7 (BD Biosciences) and anti-NKp46-APC (Miltenyi Biotec) in addition to EMA and the lineage-specific antibodies described above.

Serti E., Chepa-Lotrea X., Kim Y.J., Keane M., Fryzek N., Liang T.J., Ghany M, & Rehermann B. (2015). Successful Interferon-Free Therapy of Chronic Hepatitis C Virus Infection Normalizes Natural Killer Cell Function. Gastroenterology, 149(1), 190-200.e2.

+ Open protocol

+ Expand

CAR T Cell-Mediated Cytotoxicity Assay

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

NB target cells were stained with VPD450 (BD Horizon) and plated onto 48-well cell culture plates at 50,000 cells/well and left to adhere overnight. Suspension target cells (697 and CMK’s) were also stained with VPD450 and re-plated into T25 flasks overnight and re-counted and plated into 48-well cell culture plates at 50,000 cells/well the next day. Primary CAR T cells were added at various effector to target ratios in a total volume of 300 μL and incubated at 37°C for 4 h or 12 h. Suspension cell wells were harvested and added to flow tubes, while adherent cell suspensions were collected, and adherent cells gently taken off the wells with Accutase (Stemcell Technologies) to prevent spontaneous flipping of phosphatidylserine and added to corresponding flow tubes. Cells were washed 1x with Annexin V Binding Buffer (BioLegend), stained with 3 μL Annexin V-APC (BioLegend) and 3 μL anti-CD69-APC-Cy7 (BD Pharmingen), and washed 1x with Annexin V Binding Buffer. 7-AAD Viability Dye (BioLegend) was added 2 min before flow cytometry analysis. Detection was done by flow cytometry (Auora Cytek) and analyzed with FlowJo software (v10). Percent cytotoxicity was measured by adding Annexin V+, 7AAD+, and Annexin V+7AAD+ target cells.

Lee J.Y., Jonus H.C., Sadanand A., Branella G.M., Maximov V., Suttapitugsakul S., Schniederjan M.J., Shim J., Ho A., Parwani K.K., Fedanov A., Pilgrim A.A., Silva J.A., Schnepp R.W., Doering C.B., Wu R., Spencer H.T, & Goldsmith K.C. (2023). Identification and targeting of protein tyrosine kinase 7 (PTK7) as an immunotherapy candidate for neuroblastoma. Cell Reports Medicine, 4(6), 101091.

+ 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!