Clone hit8a

Manufactured by BD

Sourced in United States

The Clone HIT8A is a laboratory instrument used for the amplification and detection of genetic material. It is a core tool for molecular biology research and applications.

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Lab products found in correlation

Pe conjugated mabs, by BD (1 mentions) Facscanto 2, by BD (1 mentions) Facsdiva software, by BD (1 mentions)

2 protocols using clone hit8a

Quantifying CD8+ T Cell Proliferation

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EXAMPLE 8

T Cell Proliferation Assay. CD8⁺ T cell proliferation was measured using a CFSE assay as we described by Chentoufi, A et al., A novel HLA (HLA-A*0201) transgenic rabbit model for preclinical evaluation of human CD8⁺ T cell epitope-based vaccines against ocular herpes. J. Immunol. 184: 2561-2571 (2010), the content of which is hereby incorporated by reference in its entirety. Briefly, PBMCs were labeled with CFSE (2 μM) and incubated for 5 days with or without individual gB peptide (10 μg/ml). As a positive control, 2 μg/ml PHA was used to stimulate T cells for 3 days. The cells were then washed and stained with PE-conjugated mAbs specific to human CD8 molecules (clone HIT8A; BD Pharmingen). The numbers of dividing CD8⁺ T cells per 300,000 total cells were analyzed by FACS. Their absolute number was calculated using the following formula: number of events in CD8⁺/CFSE⁺ cells×number of events in gated lymphocytes/number of total events acquired.

US09878033B2. Immunogenic peptides for treatment of herpes simplex virus infection and conditions (2018-01-30). The Regents of the University of California [US]. Inventors: Lbachir BenMohamed [US], Anthony Nesburn [US].

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GAD65-Specific CD8+ T Cell Quantification

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On day 6, following previously published protocols [37 (link),38 ], approximately 5x10⁵ cells, stimulated with the GAD65 AA 114–122 peptide (supplemented with IL-2) or cultured with IL-2 alone, were washed and finally resuspended in approximately 50 μl. 1 μl of phycoerithrin (PE) labelled GAD65 AA 114–122 pentamer was added to each cell preparation and incubated in ice for 30 minutes in the dark, then washed in wash buffer. Mouse monoclonal antibody (mAb) anti-human CD8 at 1:20 dilution (fluorescein (FITC) labelled, clone HIT8a, cat# 555634, Becton & Dickinson (BD), Pharmingen, San Diego, CA, USA) and mouse mAb anti-human CD3 at 1:20 dilution (allophycocyanin (APC) labelled, clone UCHT1, cat# 555335, BD, Pharmingen) were added for further discriminating the different cell preparations. After staining, cells were immediately acquired for the analysis on the FACSCanto II (BD). Flow cytometric profiles were analyzed using the FACSDiva software (BD Bioscience). Dead cells were excluded from the analysis by side/forward scatter gating [39 (link)]. A minimum of twenty thousand events, gated on living cells, were collected per dataset.

Perri V., Gianchecchi E., Cifaldi L., Pellegrino M., Giorda E., Andreani M., Cappa M, & Fierabracci A. (2017). Identification of GAD65 AA 114-122 reactive 'memory-like' NK cells in newly diagnosed Type 1 diabetic patients by HLA-class I pentamers. PLoS ONE, 12(12), e0189615.

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