Anti cd8 apc cy7

Manufactured by BD

Sourced in United States

Anti-CD8-APC-Cy7 is a fluorochrome-conjugated monoclonal antibody that binds to the CD8 antigen expressed on the surface of cytotoxic T cells. It is designed for use in flow cytometry applications.

Automatically generated - may contain errors

Lab products found in correlation

27 protocols using anti cd8 apc cy7

Detailed Immune Cell Phenotyping

Cited 2 times

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

After homogenization of spleen tissue, immune cells were purified using 35% Percoll (GE Healthcare, La Chapelle-sur-Erdre, France) and red blood cells were lyzed. 10⁶ leucocytes were incubated with anti-CD16/32 (BD Pharmingen, Rungis, France) to block non-specific binding and washed. Cells were then incubated 30 min with appropriate dilutions of anti-CD3-Pacific Blue, anti-CD8-APC-Cy7, anti-CD4-PE, anti-NK1.1-Percp-Cy5.5 and anti-CD19-APC antibodies, all purchased from BD Pharmingen. The staining of ST2 was assessed with a rat monoclonal anti-mouse ST2-FITC antibody (clone DJ8; MB Bioproducts). Cells were washed, fixed in PBS containing 2% FCS, 0.01 M sodium azide and 2% formaldehyde and analyzed on a FACS Aria II ® flow cytometer using BD FACS Diva software (BD Biosciences). Dead cells and doublet cells were excluded on the basis of forward and side scatter. The different immune cell types were identified and gated as follows: BL were CD19⁺; NK cells were NK1.1⁺/CD3⁻; T CD8⁺ lymphocytes were NK1.1⁻/CD3⁺/CD8⁺/CD4⁻; and T CD4⁺ lymphocytes were NK1.1⁻/CD3⁺/CD4⁺/CD8⁻. The gating strategy was previously described [5 (link), 12 (link)].

Lamberet A., Rostan O., Dion S., Jan A., Guegan H., Manuel C., Samson M., Gangneux J.P, & Robert-Gangneux F. (2020). IL-33/ST2 axis is involved in disease progression in the spleen during Leishmania donovani infection. Parasites & Vectors, 13, 320.

+ Open protocol

+ Expand

Antibody Panel for Immune Cell Profiling

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

Antibodies were purchased from Santa Cruz Biotechnology (USA), BD Biosciences (USA) and Imgenex (Bhubaneswar, Odisha, India). Anti-MBP antibody (Cat No. 808) was from Santa Cruz. Anti-CD3-FITC (Cat No. 555274), anti-CD8-APC-Cy7 (Cat No. 557654), anti-CD19-APC (Cat No. 550992), anti-CD25-PECy7 (Cat No. 552880) and anti-CD45-APC (Cat No. 559864) were from BD Biosciences. Anti-CD4-PE (Cat No. 5922D) was purchased from Imgenex.

Nishana M., Nilavar N.M., Kumari R., Pandey M, & Raghavan S.C. (2017). HIV integrase inhibitor, Elvitegravir, impairs RAG functions and inhibits V(D)J recombination. Cell Death & Disease, 8(6), e2852-.

+ Open protocol

+ Expand

Imaging Flow Cytometry for Immune Cell Analysis

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

For imaging flow cytometry experiments: anti-CD8 APC-Cy7 (BD Biosciences, San Jose, CA or Tonbo Biosciences, San Diego, CA) and anti-CD56 PE-CF594 (BD Biosciences) were used for surface staining; all intracellular stains included anti-perforin D48 PE or BV421 (Biolegend, San Diego, CA) and anti-perforin δG9 FITC (Biolegend) and one of the following unconjugated antibodies: anti-CD71 (Cell Signaling Technology, Danvers, MA), anti-granzyme B (BD Biosciences), anti-rab3D (Abcam, Cambridge, MA), anti-rab4 (Pierce Antibodies, Waltham, MA), anti-rab5 (Cell Signaling Technology), anti-rab7 (Santa Cruz Biotechnology, Dallas, TX), anti-rab8 (Cell Signaling Technology), anti-rab11a (Cell Signaling Technology), anti-rab27a (Abcam), anti-rab35 (Abcam), anti-rab37 (Abcam), anti-syntaxin6 (Cell Signaling Technology), anti-syntaxin7 (R&D systems, Minneapolis, MN), anti-vti1b (Abcam), anti-VAMP3 (Abcam), anti-VAMP4 (Abcam), or anti-SNAP23 (Abcam). The unconjugated antibodies were detected using goat anti-rabbit AF647, chicken anti-goat AF647, or donkey anti-sheep AF647 secondary antibodies (Invitrogen, Carlsbad, CA). For confocal microscopy, perforin D48 FITC (Abcam), perforin δG9 AF647 (Biolegend), and goat anti-rabbit or donkey anti-sheep AF568 secondary antibodies (Invitrogen) were used, along with the selected rab or SNARE antibody.

Lesteberg K.E., Orange J.S, & Makedonas G. (2017). Recycling endosomes in human cytotoxic T lymphocytes constitute an auxiliary intracellular trafficking pathway for newly synthesized perforin. Immunologic research, 65(5), 1031-1045.

+ Open protocol

+ Expand

T Cell Activation Assay with rF1-V and SA-4-1BBL

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

4 × 10⁶ lymphocytes from peripheral lymph nodes were plated in 0.5 ml in a 48 well-plate and stimulated with rF1-V protein (20 µg) and SA-4-1BBL (25 ng) and 20 U IL-2 for 24 hours. Wells without proteins served as negative controls. 1 µl/ml Golgi Plug (BD Pharmingen) was added during the last 4 h of incubation. Cells were surface stained with anti-CD4-Alexa700 (BD Biosciences; Cat. No. 557956), anti-CD8-APC-Cy7 (BD Biosciences; Cat. No. 557654) and anti-CD44-APC (eBiosciences; Cat. No. 17–0441-83) antibodies and fixed with 4% paraformaldehyde for 15 min. Following permeabilization, cells were stained with anti-IFN-γ-PE-Cy7 (eBiosciences; Cat. No. 25–7311-82) or isotype controls (eBiosciences; Cat. No. 12–4301-82, 25–4301-82, and 17–4031-82) and analyzed using flow cytometry (BD FACS LSR-II and FACSDiva software).

Bowen W., Batra L., Pulsifer A.R., Yolcu E.S., Lawrenz M.B, & Shirwan H. (2019). Robust Th1 cellular and humoral responses generated by the Yersinia pestis rF1-V subunit vaccine formulated to contain an agonist of the CD137 pathway do not translate into increased protection against pneumonic plague. Vaccine, 37(38), 5708-5716.

+ Open protocol

+ Expand

Comprehensive T-cell Characterization Assays

Cited 1 time

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

IFN-γ ELISPOT and cytometric bead array (CBA) assays were carried out as previously described [37 (link)] and intracellular cytokine staining and staining of cell surface molecules was carried out as previously described by Burgers et al., 2006 [47 (link)]. The following antibodies were used; anti-CD3⁺ Alexa 700, anti-CD4⁺ PE-Cy7, anti-CD8⁺ APC-Cy7, anti-CD62L APC, and anti-CD44 FITC). The cytokine antibodies were all PE-conjugated (0.2μg anti-TNF-PE, 0.06 μg anti-IL-2-PE, 0.06 μg anti-IFN-γ-PE) and were obtained from BD biosciences. The anti-CD4⁺ PE-Cy7, anti-CD8⁺ APC-Cy7 were obtained from BD biosciences. All the other antibodies were obtained from eBioscience.

Jongwe T.I., Chapman R., Douglass N., Chetty S., Chege G, & Williamson A.L. (2016). HIV-1 Subtype C Mosaic Gag Expressed by BCG and MVA Elicits Persistent Effector T Cell Responses in a Prime-Boost Regimen in Mice. PLoS ONE, 11(7), e0159141.

+ Open protocol

+ Expand

Multiparametric Flow Cytometry Profiling of Immune Cells

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

The following antibodies were used for flow cytometric analysis: Anti-Perforin-Alexa fluor 488, anti-CD28-allophycocyanin (APC), anti-CD19-APC, anti-CD3-APC, anti-CD56-APC, anti-CD3-APC-cyanin7 (Cy7), anti-CD8-APC-Cy7, anti-CD14-APC-Cy7, anti-CD14-fluorescein isothiocyanate (FITC), anti-HLA-DR-FITC, anti-CD16-R-phycoerythrin (PE), anti-GATA3-PE, anti-CD45RA-PE-cyanin 5 (Cy5), anti-CD4-PE-Cy5, anti-CD16-PE-Cy5, anti-CCR7-PE-Cy7, anti-CD4-PE-Cy7, anti-CD4-V450, anti-GranzymeB-V450, anti-CD8-V500, anti-CD3-V500 (all from BD Bioscience, Franklin Lakes, NJ), anti-CD57-FITC, anti-CD4-FITC, anti-CD85j-PE, anti-CX3CR1-PE, anti-T-bet-PE-Cy7, anti-Eomes-Peridinin chlorophyll (PerCP)-efluor710 (six from eBioscience, San Diego, CA), anti-HLA-DR-PE-Cy5, anti-IL-7Rα-V450, anti-CD57-V450 (three from BioLegend, San Diego, CA), anti-CX3CR1-FITC (MBL International Corporation, Woburn, MA). For intracellular staining of T cell lineage-specific transcription factors (T-bet, Gata3, and Eomes), granzyme B and perforin, PBMC were fixed and permeabilized with Fix/Perm buffer set (BioLegend). Stained cells were acquired by a BD LSRFortessa (BD bioscience) and analyzed by using FlowJo software (ver. 9.0 or 10.0; Tree Star, OR).

Kim H.Y., Yoo T.H., Hwang Y., Lee G.H., Kim B., Jang J., Yu H.T., Kim M.C., Cho J.Y., Lee C.J., Kim H.C., Park S, & Lee W.W. (2017). Indoxyl sulfate (IS)-mediated immune dysfunction provokes endothelial damage in patients with end-stage renal disease (ESRD). Scientific Reports, 7, 3057.

+ Open protocol

+ Expand

Characterization of WT1-Specific T Cells

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

TAME was performed as previously described^{31 (link), 32 (link)} in HLA-A2.1 transgenic mice. Briefly, single-cell suspensions of pooled spleen and lymph nodes were stained with WT1A-PE, WT1B-APC or both tetramers before magnetic enrichment. Mice were randomly divided into groups of five. Enriched cells were then stained with mouse anti-CD3 PerCP-Cy5.5, anti-CD8 APC-Cy7 (#557654, BD Biosciences), anti-CD44 PE-Cy7 (#25044182, eBiosciences), anti-CD62L BV570 (#104433, Biolegend), anti-CD4 FITC (#553055, BD Biosciences), anti-B220 FITC (#553088, BD Biosciences), anti-F4/80 FITC (#11480185, eBiosciences), anti-CD11b FITC (#11011282, eBiosciences), anti-CD11c FITC (#11011482, eBiosciences), anti-I-A^b FITC (#116406, Biolegend) and LIVE/DEAD Fixable Aqua before acquiring using the LSR Fortessa II.

Nguyen T.H., Tan A.C., Xiang S.D., Goubier A., Harland K.L., Clemens E.B., Plebanski M, & Kedzierska K. (2017). Understanding CD8+ T-cell responses toward the native and alternate HLA-A*02:01-restricted WT1 epitope. Clinical & Translational Immunology, 6(3), e134-.

+ Open protocol

+ Expand

Intracellular Cytokine Profiling of Activated PBMCs

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

PBMCs were stimulated with anti-CD3 antibody (0.1 μg/ml) for 6 h. After 1 h of incubation, brefeldin A and monensin (BD Biosciences) were added to stimulate intracellular cytokine protein accumulation. Following surface staining with anti-CD3-PE-Cy7, anti-CD4-Horizon V500, anti-CD8-APC-Cy7, anti-CD31-FITC, anti-CXCR4-PerCP-Cy5.5, and anti-CD28-APC-H7, the cells were fixed and permeabilized using the Fixation/Permeabilization Buffer Kit and further stained for intracellular cytokines with anti-TNF-α-FITC, anti-IFN-γ-FITC, anti-IL-6-PE, and anti-IL-10-APC (all from BD Biosciences).

Zhang G., Liu Y., Qiu Y., Zhang J., Sun J., Zhou Z., Wang Z., Zeng P., Tao J, & He J. (2020). Circulating senescent angiogenic T cells are linked with endothelial dysfunction and systemic inflammation in hypertension. Journal of Hypertension, 39(5), 970-978.

+ Open protocol

+ Expand

Comprehensive Immune Phenotyping of Tumor Samples

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

Tumor tissue was harvested and cut into small fragments followed by digestion with a tumor disassociation kit (Miltenyi Biotec, USA) for 30 min, and then filtered by 70 μm cell strainers. Cells were stimulated with phorbol myristate acetate (PMA)/ionomyocin in the presence of Golgi stop and Monesin (eBioscience) for 4 h. Cells were washed with PBS and then stained with Live/Dead dye and fluorochrome-labeled antibodies specific to cell surface markers [anti-CD45 Alexa 700 (Clone 30-F11, BioLegend), anti-PD-L1 PE-Cy7 (Clone 10F.9G2, BioLegend), anti-CD3 PerCP (Clone 45-2C11, BioLegend), anti-CD8 APC-Cy7 (Clone 53–6.7, BD), anti-CD11c FITC (ab210308, Abcam), anti-PD1 PE (Clone RMP1–30, BioLegend), anti-CD4 PE-CF594 (Clone RM4–5, BD), anti-MHC-I PE (Clone 28–14-8, BioLegend), anti-CD44 APC-Cy7 (Clone IM7, BioLegend), anti-CD62L APC (Clone MEL-14, BioLegend), anti-CD25 PerCP (Clone 3C7, BioLegend), anti-CD127 FITC (Clone A7R34, BioLegend), anti-ICOS PE-Cy7 (Clone 7E.17G9, BioLegend)]. After fixation–permeabilization, cells were stained with fluoro-conjugated antibodies specific to intracellular markers [anti-Ki67 APC (Clone 16A8, BioLegend), anti-IFNγ APC (Clone XMG1.2, BioLegend), anti-granzyme B FITC (Clone GB11, BioLegend)] or with the isotype control. Flow cytometry analysis was performed on BD LSRFortessa and data were analyzed by FlowJo V10.

Hong S., Bi M., Yu H., Yan Z, & Wang H. (2020). Radiation therapy enhanced therapeutic efficacy of anti-PD1 against gastric cancer. Journal of Radiation Research, 61(6), 851-859.

+ Open protocol

+ Expand

Cytokine Profile of Antigen-Specific T Cells

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

1.0 × 10⁶ PBMCs were stimulated with 1 μg/ml mesothelin peptide pool (Peptides & Elephants, Potsdam, Germany), medium control or PMA/Ionomycin (positive control, Sigma-Aldrich, St Louis, MI; USA) in R10 medium in the presence of Brefeldin A (10μg/mL, Sigma-Aldrich St Louis, MI, USA) for 6 hours at 37°C. Stimulation was stopped by transferring the cells to a 4°C refrigerator, followed by washing with FACS buffer and staining with the following reagents: anti-CD3 Pacific blue (BD Biosciences, CA, USA), anti-CD4 PerCP-Cy5.5 (BD Biosciences, CA, USA) and anti-CD8 APC-Cy7 (BD Biosciences, CA, USA). After a 15-minute incubation at 4°C, the cells were washed and fixed with a Fix/Perm reagent (Beckman coulter, CA, USA), followed by a further 30-minute incubation at 4°C with an intracellular antibody mix (anti-TNFα APC (BD Biosciences CA, USA), anti-IFN-γ PE-Cy7 (BD Biosciences CA, USA), anti-IL-2 PE (BD biosciences CA, USA) and anti-IL-17 FITC (BioLegend, CA, USA)). The stained cells were then washed with FACS buffer and acquired on a FACSAria flow cytometer (BD Biosciences, Stockholm, Sweden). Data was analysed using FlowJo software (Treestar Inc.).

Liu Z., Rao M., Poiret T., Nava S., Meng Q., von Landenberg A., Bartek J J.r., Xie S., Sinclair G., Peredo I., Dodoo E, & Maeurer M. (2017). Mesothelin as a novel biomarker and immunotherapeutic target in human glioblastoma. Oncotarget, 8(46), 80208-80222.

+ 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!