Anti cd107a pe antibody

Manufactured by BioLegend

Sourced in United States

The Anti-CD107a-PE antibody is a fluorescent-labeled monoclonal antibody that binds to the CD107a (LAMP-1) cell surface protein. CD107a is a marker for degranulation, a process that occurs during the activation of cytotoxic T cells and natural killer cells. The PE (phycoerythrin) fluorescent dye is conjugated to the antibody, allowing for the detection and analysis of CD107a-positive cells using flow cytometry.

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Lab products found in correlation

Pkh26, by Merck Group (1 mentions) Pi stain, by Merck Group (1 mentions) Monensin, by BioLegend (1 mentions) Monensin, by Merck Group (1 mentions) Anti cd3 percp cy5, by BioLegend (1 mentions) Anti cd8 apc cy7, by BioLegend (1 mentions) Anti cd4 pe cy7, by BioLegend (1 mentions) Facs canto 1 cytometer, by BD (1 mentions) Cytofix cytoperm permeabilization kit, by BD (1 mentions) Brefeldin a, by Merck Group (1 mentions) Facsdiva software, by BD (1 mentions) Golgistop, by BD (1 mentions) Ril 15, by R&D Systems (1 mentions) Ril 12, by R&D Systems (1 mentions) Lsrii flow cytometer, by BD (1 mentions) Facsaria, by BD (1 mentions)

Close spelling variants of this product

PE-conjugated anti-CD107a antibody Anti-CD107a-PE Anti-CD107a antibody CD107a-PE Anti-CD107a

6 protocols using anti cd107a pe antibody

Quantifying CAR-T Cell Proliferation and Cytotoxicity

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Proliferation of T cells was determined by PKH26 staining in vitro. Individual groups of CAR-T and control Utd T cells (1 × 10⁶ cells/tube) were labeled with PKH26 (Sigma-Aldrich) at 37 °C for 5 min. After being washed, individual groups of cells were stimulated with the same number of CD105⁺ target cells that had been irradiated (100 Gy) for 120 h. The suspended T cells were collected and the percentages of proliferative T cells were determined by flow cytometry.
For cytotoxicity assays, individual groups of CAR-T and control Utd T cells were cultured in triplicate with each type of PKH26-labeled target cells at a ratio of 3 : 1, 1 : 1, or 1 : 3 for 16 h. After being washed, the adhered target cells were collected and stained with PI stain (Sigma), and the percentages of PKH26⁺PI⁺ dead cells in individual groups were determined by flow cytometry as % of specific lysis. In addition, anti-CD105 CAR-T cells were co-cultured with Bel7404 cells at a ratio of 3 : 1 for 16 h in the presence or absence of CD105 protein (eBioscience, Cat14-1051-85). The percentages of lysed target cells were determined by flow cytometry. Moreover, individual groups of CAR-T and control Utd T cells were cultured in triplicate with Bel7404 cells at a ratio of 3 : 1 for 12 h and stained with PE-anti-CD107a antibody (Biolegend,Cat304124). The percentages of CD107a⁺ T cells were determined by flow cytometry.

Mo F., Duan S., Jiang X., Yang X., Hou X., Shi W., Carlos C.J., Liu A., Yin S., Wang W., Yao H., Yu Z., Tang Z., Xie S., Ding Z., Zhao X., Hammock B.D, & Lu X. (2021). Nanobody-based chimeric antigen receptor T cells designed by CRISPR/Cas9 technology for solid tumor immunotherapy. Signal Transduction and Targeted Therapy, 6, 80.

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CD8+ T Cell Degranulation Assay

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The CTLs from each group were co-cultured with the target cells at an E:T ratio of 10:1. The PE anti-CD107a antibody (Biolegend, USA) was included during the culture. After 1 h, 2μM monensin (Biolegend, USA) was added to the co-cultured system and incubated for an additional 4 h, and then the cells were stained with the APC anti-human CD8a and analyzed using ow cytometry. All the CD8+ CD107a+ cells were regarded as the degranulated T cells.

Li Z., Chen X., Liu L., Zhou M., Zhou G, & Liu T. (2022). Development of the ^T-ALLiPSC-based therapeutic cancer vaccines for T-cell acute lymphoblastic leukemia. Medical oncology (Northwood, London, England), 39(12).

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NK Cell Cytotoxicity Assay Using CD107a

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NK cell cytotoxic activity was assessed by CD107a degranulation. Briefly, after overnight stimulation with recombinant human IL-12 (10 ng/mL) and IL-18 (50 ng/mL), or only culture medium, PBMCs were incubated for 3 h at 37 °C with K562 target cells (E:T = 5:1) in the presence of anti-CD107a-PE antibody (Biolegend, CA, United States) and monensin (2 µM). Cells were then labeled with CD3-PerCP/Cy5.5 and CD56-FITC before flow cytometry analysis.

Wang W.T., Zhao X.Q., Li G.P., Chen Y.Z., Wang L., Han M.F., Li W.N., Chen T., Chen G., Xu D., Ning Q, & Zhao X.P. (2019). Immune response pattern varies with the natural history of chronic hepatitis B. World Journal of Gastroenterology, 25(16), 1950-1963.

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Evaluation of TCL Reactivity by Flow Cytometry

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On day 7 to 14 after primary stimulation, reactivity of TCL was examined by flow cytometry. Cells were washed three times and incubated in the presence or absence of 10 µg/ml abacavir for 6 h with CFSE labelled autologous PBMC. After 2 h of co-incubation Brefeldin A (10 µg/ml, Sigma-Chemicals, Buchs, Switzerland), Monensin (6 µg/ml, Sigma-Chemicals, Buchs, Switzerland) and anti-CD107a-PE antibody (Biolegend, San Diego, CA, USA) were added. Surface staining was performed with anti-CD3-PerCp-Cy5.5, anti-CD4-PE-Cy7 and anti-CD8-APC-Cy7 (Biolegend, San Diego, California). Intracellular staining with anti-IFNγ-APC was performed according to the Cytofix/Cytoperm permeabilization kit (BD Biosciences, Basel, Switzerland). FACS analysis was performed on a FACSCanto-I cytometer using FACS-Diva software (BD Biosciences).

Adam J., Wuillemin N., Watkins S., Jamin H., Eriksson K.K., Villiger P., Fontana S., Pichler W.J, & Yerly D. (2014). Abacavir Induced T Cell Reactivity from Drug Naïve Individuals Shares Features of Allo-Immune Responses. PLoS ONE, 9(4), e95339.

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Cytokine Modulation of PBMC Function

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PBMCs were co-cultured with different concentrations of rIL-10 and/or rTGF-β for 5 h, then stimulated with rIL-12 (10 ng/ml, R&D) and rIL-15 (50 ng/ml, R&D) for 24 h at 37 °C, and anti-CD107a-PE antibody (Biolegend, San Diego, CA, USA) added simultaneously. Cells were co-cultured in GolgiStop™ ((Becton Dickinson) for ≥5 h before the end of stimulation. PBMCs were collected, and anti-CD3-Percp and anti-CD56-PE-cy7 were used for surface staining. Cells were then stained for intracellular IFN-γ with anti-IFN-γ-APC antibody (BD Biosciences, San Jose, CA, USA), washed with phosphate-buffered saline, and fixed in 1% paraformaldehyde, followed by analysis using the LSR II flow cytometer.

Jiang Y., Yang M., Sun X., Chen X., Ma M., Yin X., Qian S., Zhang Z., Fu Y., Liu J., Han X., Xu J, & Shang H. (2018). IL-10+ NK and TGF-β+ NK cells play negative regulatory roles in HIV infection. BMC Infectious Diseases, 18, 80.

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CRISPR-Based T-Cell Cytotoxicity Assay

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mm10dgLib lentivirus-infected OT-I;Cas9β CD8⁺ T cells were cultured on the 10-cm dishes pretreated with anti-CD3ε in the cRPMI-1640 supplemented with 2 ng / mL IL-2, 1 μg / mL anti-CD28, and 2 ng / mL IL-12p70 for 4 days, the media were changed with fresh media every day. About 12 hours before the kill assay, infected OT-I;Cas9β CD8⁺ T cells were reseeded onto new dishes without treatment with anti-CD3ε antibody, and cultured with cRPMI only supplemented with 2 ng / mL IL-2 and 2 ng / mL IL-12p70 to rest cells. At the same time, 2e6 E0771 cells were seeded in 6-well plates in D10 media. On day 5, E0771 cells were incubated with 1 ng / mL SIINFEKL peptide for 4 hours. Before start kill assay, CD8⁺ T cells were suspended with fresh cRPMI media supplied with 2 nM monensin and anti-CD107a-PE antibody (BioLegend) (1:400 dilution), the final cell concentration was 2e6 cells / mL. After SIINFEKL peptide incubation, 3e6 CD8⁺ T cells per replicate (E0771: T cell = 1:1) were added into E0771 cells for 2 h co-culture. At the end of co-culture, T cells were gently washed down with PBS and stained with anti-CD8α-APC, anti-CD3ε-PE/Cy7 for 30 min on ice, cells were analyzed and sorted using BD FACSAria. CD107a-high cells were sorted by FACS for library readout similar to an in vitro T cell cytotoxicity CRISPR screen approach in a previous study (Dong et al., 2019 (link)).

Ye L., Park J.J., Peng L., Yang Q., Chow R.D., Dong M.B., Lam S.Z., Guo J., Tang E., Zhang Y., Wang G., Dai X., Du Y., Kim H.R., Cao H., Errami Y., Clark P., Bersenev A., Montgomery R.R, & Chen S. (2022). A genome-scale gain-of-function CRISPR screen in CD8 T cells identifies proline metabolism as a means to enhance CAR-T therapy. Cell metabolism, 34(4), 595-614.e14.

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