Anti cd107a pe

Manufactured by BioLegend

Anti-CD107a-PE is a monoclonal antibody that binds to the CD107a (LAMP-1) antigen expressed on the surface of cells. CD107a is a marker of lysosomal degranulation and is used to detect the activation of cytotoxic cells, such as natural killer (NK) cells and cytotoxic T lymphocytes (CTLs).

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Lab products found in correlation

Golgiplug, by BD (2 mentions) Anti tnf apc, by BioLegend (1 mentions) Ionomycin, by Merck Group (1 mentions) Anti ifn γ fitc, by BioLegend (1 mentions) Efluor 670, by Thermo Fisher Scientific (1 mentions) Anti cd8a apc cy7, by BioLegend (1 mentions) Clone h4a3, by BioLegend (1 mentions) Permeabilization solution, by BD (1 mentions) Anti ifn γ fitc, by BD (1 mentions) Facscanto 2, by BD (1 mentions) Anti cd3 anti cd28 dynabeads, by Thermo Fisher Scientific (1 mentions) Anti cd45 fitc, by Thermo Fisher Scientific (1 mentions) Anti cd69 pe, by BioLegend (1 mentions) Anti cd8 apc, by BioLegend (1 mentions) Dnase, by Merck Group (1 mentions) Collagenase type 4, by Merck Group (1 mentions) Live dead fixable aqua dead cell stain, by Thermo Fisher Scientific (1 mentions) Lsrii flow cytometer, by BD (1 mentions)

Close spelling variants of this product

Anti-CD107a (26 citations) CD107a-PE (8 citations) Anti-CD107a-PE-Cy7 (8 citations) PE-conjugated anti-CD107a antibody (8 citations) Anti-CD107a-PE antibody (6 citations) PE anti-human CD107a (4 citations) Anti-CD107a PE (ID4B, (2 citations) Anti-CD107a-PE/Cy7 (H4A3) (2 citations) Anti-CD107a-PE (clone H4A3, (2 citations) Anti-CD56-PE and anti-CD107a-APC antibodies (1 citations)

6 protocols using anti cd107a pe

TCR-Dependent Activation of PBMCs

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Peripheral blood mononuclear cells were cultured with anti-CD107a PE (Biolegend), monensin, brefeldin A, phorbol 12-myristate 13-acetate (PMA), and ionomycin (Sigma) for 4 h, then stained with anti-IFNγ FITC, anti-TNF APC, and anti-IL-2 APC-Cy7 (Biolegend or eBioscience), and analyzed by flow cytometry.
To evaluate the effect of TCR-dependent activation, PBMCs were cultured with anti-CD3 (1 µg/mL) and anti-CD28 (0.5 mg/mL). After 72 h, PBMCs were cultured with monensin, brefeldin A, and anti-CD107a PE for 6 h and then stained with anti-human CD8 APCCy7, anti-NT rabbit and anti-rabbit Alexa 647, anti-IL-2 PECy7, anti-TNF PerCPCy5.5, or anti-IFNγ PerCPCy5.5.

Sanmarco L.M., Visconti L.M., Eberhardt N., Ramello M.C., Ponce N.E., Spitale N.B., Vozza M.L., Bernardi G.A., Gea S., Minguez A.R, & Aoki M.P. (2016). IL-6 Improves the Nitric Oxide-Induced Cytotoxic CD8+ T Cell Dysfunction in Human Chagas Disease. Frontiers in Immunology, 7, 626.

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CTL Cytotoxicity Assay with P815 Targets

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CTLs obtained from PBMC at day 7 post stimulation and day 2 after purification were mixed at a 1:1 ratio with P815 target cells previously stained with cell proliferation dye efluor 670 (1/2000; ebiosciences) and coated with indicated concentration of anti-CD3 (OKT3 produced by A. Alcover) in the presence of anti-CD107a-PE (1/90, clone H4A3, Biolegend) and incubated for 3h at 37°C. Cells were stained with anti-CD8a-APC-Cy7 (1/30; clone HI100; Biolegend). Facs analyses were performed as above.

Cuche C., Mastrogiovanni M., Juzans M., Laude H., Ungeheuer M.N., Krentzel D., Gariboldi M.I., Scott-Algara D., Madec M., Goyard S., Floch C., Chauveau-Le Friec G., Lafaye P., Renaudat C., Le Bidan M., Micallef C., Schmutz S., Mella S., Novault S., Hasan M., Duffy D., Di Bartolo V, & Alcover A. (2023). T cell migration and effector function differences in familial adenomatous polyposis patients with APC gene mutations. Frontiers in Immunology, 14, 1163466.

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Quantifying T-cell Activation and Cytokine Production

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Fixation/permeabilization solution Kit with BD GolgiPlug (BD) was used according to the manufacturer’s instructions. Briefly, T2-A24 cells were pulsed with 20 µM of the indicated synthetic peptides for 1.5 h at 25°C and served as antigen-presenting cells. The indicated T-cell clones were incubated with T2-A24 cells at a 2:1 ratio for 4 h at 37 °C in the presence of anti-CD107a-PE (BioLegend), GolgiPlug, and human FcR blocking reagent (Clear Back, MBL). The cells were stained with anti-CD8 PC5, followed by permeabilization using a permeabilization solution (BD). The cells were subsequently stained with anti-IFNγ FITC and analyzed using FACS Canto II (BD).

Shinkawa T., Tokita S., Nakatsugawa M., Kikuchi Y., Kanaseki T, & Torigoe T. (2021). Characterization of CD8+ T-cell responses to non-anchor-type HLA class I neoantigens with single amino-acid substitutions. Oncoimmunology, 10(1), 1870062.

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Assessing Cargo Release in Activated CTLs

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NC-CTLs loaded with Alexa647-OVA were co-cultured for 3 – 16 hours (as indicated in figure legends) with anti-CD3/anti-CD28 Dynabeads (Thermo Fisher Scientific) at the indicated NC-CTL:beads ratios; or with an excess of BLCLs matched on the restricting MHC-I allele and pulse/washed with cognate peptide. Co-cultures were performed in R10–50 medium at 37°C. In some experiments supernatants were retained to quantify released cargo by ELISA. Cells were then stained with fluorochrome-conjugated Abs to CD3 and CD8 (Biolegend). Cargo release was assessed by flow cytometry, gating on CD3⁺CD8⁺ lymphocytes and assessing cell-associated cargo by Alexa647 fluorescence. In some experiments, we simultaneously assessed degranulation (CD107a exposure) by adding anti-CD107a PE (Biolegend) at the beginning of the co-culture period. Note that Brefeldin A was not used in these experiments. To test the effects of the perforin inhibitor concanamycin A (CMA) on triggered release, NC-CTLs were co-cultured with the indicated concentrations of CMA (Sigma) for 3 hours, washed three times, then tested in triggered release assays as above.

Jones R.B., Mueller S., Kumari S., Vrbanac V., Genel S., Tager A., Allen T., Walker B.D, & Irvine D.J. (2016). Antigen recognition-triggered drug delivery mediated by nanocapsule-functionalized cytotoxic T-cells. Biomaterials, 117, 44-53.

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Tumor-Infiltrating Lymphocyte Isolation

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To analyze TILs, the tumor tissues were sliced into small fragments and subsequently subjected to digestion for a duration of 2.5 h using 0.6 ku/ml DNAse (D5025; Sigma) and 1 mg/ml collagenase type IV (C5138; Sigma). The cells were stained using the following antibodies: anti-CD45-FITC (11–0451-82; eBioscience; 1:100), anti-CD107a-PE (121612; Biolegend; 1:100), anti-CD8-APC (100721; Biolegend; 1:50) and anti-CD69-PE (104508; Biolegend; 1:100). All flow data were analyzed using FlowJo X.

Qin G., Bai F., Hu H., Zhang J., Zhan W., Wu Z., Li J., Fu Y, & Deng Y. (2024). Targeting the NAT10/NPM1 axis abrogates PD-L1 expression and improves the response to immune checkpoint blockade therapy. Molecular Medicine, 30, 13.

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Assessing Cytokine Production in Immunized Mice

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Cytokine production by splenocytes from immunized mice was assessed by intracellular cytokine staining as described previously.³⁴ (link) Briefly, splenocytes were stimulated with tHIVconsvX-derived 15/11 peptides assembled into 10 pools, P1–P10, for 90 min at 37°C and then for an additional 5 hr in the presence of brefeldin A (Golgiplug; BD Biosciences) to prevent cytokine secretion. Cells were surface stained with anti-CD8-R-phycoerythrin (PE)-Cy5 red (eBioscience) antibodies and LIVE/DEAD fixable aqua dead cell stain (Invitrogen) and then permeabilized and incubated with various combinations of anti-IL-2-fluorescein isothiocyanate (FITC), anti-CD107a-PE, anti-TNF-α-antigen-presenting cell (APC), and anti-IFN-γ-V450 monoclonal antibodies (BioLegend). Samples were acquired on an LSR II flow cytometer (BD Biosciences) and data were analyzed using FlowJo software (version 9.5.2; Tree Star). Plurifunctionality was assessed using Simplified Presentation of Incredibly Complex Evaluations (SPICE) software (National Institute of Allergy and Infectious Diseases [NIAID]).

Wee E.G., Ondondo B., Berglund P., Archer J., McMichael A.J., Baltimore D., ter Meulen J.H, & Hanke T. (2017). HIV-1 Conserved Mosaics Delivered by Regimens with Integration-Deficient DC-Targeting Lentiviral Vector Induce Robust T Cells. Molecular Therapy, 25(2), 494-503.

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