After centrifugation, PBMCs (2 × 105 per well) in Falcon 96‐well U bottom plates (Corning, USA) were resuspended in 20 μl mouse hybridoma supernatant for 30 min. The negative control was FACS buffer (PBS with 5% heat inactivated Gibco goat serum, Thermo Fisher Scientific), 1% BSA (Sigma‐Aldrich) and 0.01% w/v sodium azide (Sigma). All incubations were on ice in the dark. Cells washed twice with FACS buffer were resuspended in anti‐mouse Ig‐BV421 in FACS buffer and incubated for 30 min before washing with FACS buffer then PBS and re‐suspension in 1% formaldehyde in PBS. Twenty minutes later cells were washed twice and resuspended in FACS buffer for acquisition on a FACSCanto II using FACSDiva software (BD) and analysis using FlowJo software (BD).
Falcon 96 well u bottom plates
Manufactured by Corning
Sourced in United States, Germany
Falcon 96-well U-bottom plates are a type of laboratory equipment used for various cell culture and assay applications. They feature a U-shaped well bottom design and are made of high-quality materials to provide consistent results. The plates are available in a range of sizes and configurations to suit different experimental needs.
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Lab products found in correlation
2 protocols using falcon 96 well u bottom plates
1
Immunophenotyping of Mouse PBMCs
2
Amnis ImageStreamX Flow Cytometry Acquisition
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All samples were measured using the built-in autosampler from Falcon 96-well U-bottom plates (category 353077; Corning, Kaiserslautern, Germany), with 5-min acquisition time per well, on the Amnis ImageStreamX Mark II flow cytometer (Luminex Corporation, Seattle, WA, USA). All data were acquired at £60 magnification at low flow rate (0.3795 § 0.0003 mL/min, directly determined by the system) and with the removed beads option deactivated as described previously [20, 21] . The data were analyzed as described previously [22] . Additional settings can be found in supplementary Tables 2 and3.
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