Carrier and reference samples composed of equivalent amounts of untreated and LPS-stimulated murine BMDMs were prepared following the SCoPE2 protocol29 (link),33 (link), such that the carrier was composed of ~200 cells and the reference was composed of approximately five cells. This sample design was then used in the preparation of single-cell sets by mPOP50 (link), in which single cells from each condition (untreated and treated with LPS for 24 h) were sorted into a 384-well plate (Thermo, AB1384) with the cellenONE liquid-handling system (Scienion). The mixed carrier and reference sample was also used in the generation of retention-time estimate runs for the set of ten samples analyzed by pSCoPE.
Ab1384
Manufactured by Thermo Fisher Scientific
The AB1384 is a laboratory instrument designed for DNA sequencing and analysis. It is a highly capable and reliable system that leverages advanced technologies to deliver accurate and reproducible results. The core function of the AB1384 is to perform nucleic acid sequencing, a fundamental process in genomic research and genetic analysis.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using ab1384
1
scSCoPE2 protocol for single-cell analysis
2
Single-cell Mass Spectrometry Proteomics
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Carrier and reference samples composed of equivalent amounts of untreated and LPS-stimulated murine BMDMs were prepared following the SCoPE2 protocol29 (link),33 (link), such that the carrier was composed of ~200 cells and the reference was composed of approximately five cells. This sample design was then used in the preparation of single-cell sets by mPOP50 (link), in which single cells from each condition (untreated and treated with LPS for 24 h) were sorted into a 384-well plate (Thermo, AB1384) with the cellenONE liquid-handling system (Scienion). The mixed carrier and reference sample was also used in the generation of retention-time estimate runs for the set of ten samples analyzed by pSCoPE.
3
Single-cell Proteomics Sample Preparation
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Cells were harvested as single-cell suspension and prepared for MS analysis using Nano-ProteOmic sample Preparation (nPOP) as described by Leduc et al.50 ,51 . The automated collection of prepared samples had not been developed yet26 (link) and so samples were manually collected using a pipette (using 5μl of mass spectrometry grade Acetonitrile then water respectively) and transferred into a 384-well plate (Thermo AB1384). The samples were then dried down in a SpeedVac vacuum evaporator and resuspended in 1.07μl of 0.1% Formic Acid (buffer A) and tightly sealed using an aluminium foil cover (Thermo Fisher AB0626).
For the bulk experiments, cells were harvested (in MS grade water, at roughly 2000 cell/μl) and frozen at −80C. The samples were prepared using mPoP52 , following guidelines for the digest of carriers as outlined in Petelski et al.30 (link). Post digest, the samples were dried down in a SpeedVac vacuum evaporator and resuspended at a concentration of 1 μg/μl in 0.1% Formic Acid (buffer A) in a glass insert with polyspring within an HPLC vial.
For the bulk experiments, cells were harvested (in MS grade water, at roughly 2000 cell/μl) and frozen at −80C. The samples were prepared using mPoP52 , following guidelines for the digest of carriers as outlined in Petelski et al.30 (link). Post digest, the samples were dried down in a SpeedVac vacuum evaporator and resuspended at a concentration of 1 μg/μl in 0.1% Formic Acid (buffer A) in a glass insert with polyspring within an HPLC vial.
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