Conjunctivae were excised, chopped with scissors into tiny pieces, and incubated with 0.1% type IV Collagenase for 1 hour to yield single-cell suspensions. Samples were incubated with anti-CD16/32 (2.4G2, Catalog no. 553141, BD Pharmingen™, San Diego, CA), for 5 minutes at room temperature and subsequently stained with anti-CD45 (clone 30-F11, Catalog no. 103138, BioLegend) and with an infra-red fluorescent viability dye (Catalog no. L10119, Life Technologies, Grand Island, NY). The gating strategy was as follows: lymphocytes were identified by forward -scatter area (FSC-A) and side scatter area (SSC-A) gates, followed by two singlets gates (FSC-A vs. FSC-W and SSC-A vs. SSC-W) followed by live/dead identification using the infra-red fluorescent viability dye. The CD45+ cells were sorted using the Aria-II cell sorter at the Baylor College of Medicine cytometry and cell sorting core.
Catalog no l10119
Manufactured by Thermo Fisher Scientific
The Catalog no. L10119 is a laboratory equipment item manufactured by Thermo Fisher Scientific. It is a specialized piece of apparatus designed for use in scientific research and analysis. The core function of this product is to perform specific tasks within a laboratory setting, though a detailed description of its intended use cannot be provided without the risk of bias or extrapolation.
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Lab products found in correlation
3 protocols using catalog no l10119
1
Isolation of Immune Cells from Conjunctiva
2
Isolation of Mouse Corneal Immune Cells
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Adult mouse corneas anterior to the limbus were excised and incubated in 20 mM EDTA-PBS for 20 min, a total of 50 corneas from 25 mice were pooled and chopped with scissors into tiny pieces and incubated with 0.1% type IV Collagenase for 1 h to yield single-cell suspensions. The pooled samples were incubated with anti-CD16/32 (2.4G2, Catalog no. 553141, BD Pharmingen™, San Diego, CA), for 5 min at room temperature and subsequently stained with anti-CD45 (clone 30-F11, Catalog no. 103138, BioLegend) and with an infra-red fluorescent viability dye (Catalog no. L10119, Life Technologies, Grand Island, NY). The gating strategy was as follows: lymphocytes were identified by forward-scatter area (FSC-A) and side scatter area (SSC-A) gates, followed by two singlets gates (FSC-A vs. FSC-W and SSC-A vs. SSC-W) followed by live/dead identification using the infra-red fluorescent viability dye. The CD45+cells were sorted using the Aria-II cell sorter at the Baylor College of Medicine cytometry and cell sorting core.
3
Isolation of Immune Cells from Conjunctiva
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Conjunctivae were excised, chopped with scissors into tiny pieces, and incubated with 0.1% type IV Collagenase for 1 hour to yield single-cell suspensions. Samples were incubated with anti-CD16/32 (2.4G2, Catalog no. 553141, BD Pharmingen™, San Diego, CA), for 5 minutes at room temperature and subsequently stained with anti-CD45 (clone 30-F11, Catalog no. 103138, BioLegend) and with an infra-red fluorescent viability dye (Catalog no. L10119, Life Technologies, Grand Island, NY). The gating strategy was as follows: lymphocytes were identified by forward -scatter area (FSC-A) and side scatter area (SSC-A) gates, followed by two singlets gates (FSC-A vs. FSC-W and SSC-A vs. SSC-W) followed by live/dead identification using the infra-red fluorescent viability dye. The CD45+ cells were sorted using the Aria-II cell sorter at the Baylor College of Medicine cytometry and cell sorting core.
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