Project description:Halo purification samples analyzed using Multidimentional Protein identification Technology (MudPIT) on Orbitrap Velos Elite. Baits are Halo-WDR76, Halo-SPIN1, Halo-CBX5. The bait for Control Purification is Halo tag only. Purification for each bait was performed using whole cell extract from a stable cell line in 293FRT cells. 3 biological replicates for each type of bait.
Project description:Halo purification samples analyzed using Multidimentional Protein identification Technology (MudPIT) on Orbitrap Velos Elite. Baits Halo-WDR76. Purification was performed using whole cell extract from a stable cell line of 293FRT cells in WDR76 KO background. Control purification was performed on blank WDR76 KO293FRT cells. 3 biological replicates of control and Halo-WDR76.
Project description:Serial Capture Affinity Purification (SCAP) samples analyzed using Multidimentional Protein identification Technology (MudPIT) on Orbitrap Velos Elite. Baits are SNAP-SPIN1 and Halo-WDR76. SNAP purification was performed first, then Halo purification was performed. SCAP was performed using whole cell extract from a stable cell line of 293FRT cells in WDR76 KO background. 3 biological replicates of E1, UB2 and E2 fractions.
Project description:We have developed Halo-seq, an RNA proximity labeling method that allows the quantification of subcellular transcriptomes. We have demonstrated the efficacy of Halo-seq here by using it to quantify chromatin-proximal, nucleolar, and cytoplasmic transcriptomes. In Halo-seq, RNA molecules in close proximity to a spatially restricted protein are specifically marked and biotinylated, facilitating their separation from bulk cellular RNA and their quantification.
Project description:We determined the genome-wide binding profile of CTCF and cohesin subunits Smc1a and Smc3 in wild type mouse ES cells and compared them to a double FLAG-Halo-mCTCF/mRad21-SNAPf-V5 knock-in mES line, generated by CRISPR/Cas9-mediated genome editing to perform imaging experiments.
Project description:Halo purified WDR76 from a stable cell line of 293FRT in WDR76 KO background. DSSO crosslinking performed in eluted samples. Data collected using ms1/ms2/ms3 method. Data searched using Proteomics Discoverer 2.4 with XlinkX.