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XL-MS analysis of protein isolates from Flp-In 293 cells stably expressing SIN3A-HaloTag using DSSO crosslinker.

ABSTRACT: XL-MS analysis of protein isolates from Flp-In 293 cells stably expressing SIN3A-HaloTag using DSSO crosslinker.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Michael P. Washburn

PROVIDER: MSV000085083 | MassIVE | Wed Mar 11 11:28:00 GMT 2020

REPOSITORIES: MassIVE

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Title: XL-MS analysis of protein isolates from Flp-In 293 cells stably expressing SIN3B isoform 2-HaloTag using DSSO crosslinker.

Project description:XL-MS analysis of protein isolates from Flp-In 293 cells stably expressing SIN3B isoform 2-HaloTag using DSSO crosslinker.

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Project description:In effort to develop methodology for targeted top down mass spectrometry of NF kappa B p65 from human cells, we evaluated the utility of HaloTag for purification and analysis of recombinant protein. During our study, two datasets of bottom up LC-MS/MS were generated: one from in-gel digestion of the predominant band following p65-HaloTag purification, another from in-solution digestion of all the proteins present in a p65-HaloTag purification. p65-HaloTag copurifying proteins identified in our datasets include the known interactors c-Rel, NF-kappaB p105, NF-kappaB p100, and NF-kappaB inhibitor beta. Over 100 proteins were identified by at least two peptides using a Mascot ion cut-off score of 30.

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