Project description:We extracted mitochondria from Hela cells which were treated with CCCP or DMSO for 2 h. The samples were digested by FAST and peptides were used DIA to analysis.
Project description:We extracted DIP from placanta tissue of human which were diagnose preeclampsia or normal. The samples of DIPs were digested by FAST and the peptides were used DIA to analysis.
Project description:In an attempt to identify proteins interacting with transcription factor E2F2, proteins extracted from HA-E2F2 overexpressing HEK293T cells were were subjected to immunoprecipitation (IP) with an anti-HA antibody or, as a negative control (NegCt), with an anti-T antibody. Immunoprecipitated proteins were eluted, digested with trypsin, and the resulting peptides were analyzed by LC-MS/MS. The whole procedure was repeated several times in an attempt to gauge reproducibility (nine replicates of the anti-HA IP and six replicates of the control IP).
Project description:In an attempt to identify proteins interacting with transcription factor E2F2, proteins extracted from HA-E2F2 overexpressing HEK293T cells were were subjected to immunoprecipitation (IP) with an anti-HA antibody or, as a negative control (NegCt), with an anti-T antibody. Immunoprecipitated proteins were eluted, digested with trypsin, and the resulting peptides were analyzed by LC-MS/MS. The whole procedure was repeated several times in an attempt to gauge reproducibility (nine replicates of the anti-HA IP and six replicates of the control IP).
Project description:To specifically explore such mRNA localization events, we immunopurified the entire Nuclear Pore Complex scaffold using Nup59 as a bait and systematically analyzed all co-isolating mRNAs using Agilent microarrays in the budding yeast Saccharomyces cerevisiae. A mock IP was performed as a control experiment, using a yeast strain in which Nup59 was untagged. Two replicates of the Nup59 IP and of the mock IP were performed. The mRNAs with a significant enrichment (p<0.05 in the bayesian test of the LIMMA package) in the mock IP were removed from the further analyses of the Nup59-IP.
Project description:Antibodies are used in multiple cell biology applications but there are methods to assess antibody quality, riskingdata integrity and repr developed a standard operating procedure using HEK293 cells to score and antibodies for suitability in immunoprecipitation. This method, validated in in fourindependent laboratories, was developed using 1124 novel recomb for152 chromatin-related human proteins. We used mass spectrometry abundanceof all the proteinsin eachimmunoprecipitateand compared spectral abundance factors from the target antigen with those of all Antibodies for which the targetantigen or a member of its known protein most abundant protein were classified as “IP gold standard”. This m quantitative outputs that can be stored and archived in public databases, step towards a platform for community benchmarking of antibody quality.
Project description:Paired-end sequencing study of nucleosomes and immuno-precipitated Tfc1 and Brf1 complexes from MNase-digested nuclei. Nucleosomal DNA, input DNA and DNA from immunopurified TFIIIB and TFIIIC complexes (IP) were subjected to paired-end sequencing.