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Immunoprecipitation followed by mass spectrometry (IP-MS)

ABSTRACT: Immunoprecipitation of ITGB1 and YWHAE was done to understand its interactive partners in K562/S cells. Mass spectrometry was performed to identify interactive molecules. Non-specific proteins (contaminants) such as keratins were excluded from the final analysis based on the results of the negative control IgG

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Suspension Culture

SUBMITTER: Neha Agrawal

LAB HEAD: Dr. Rukmini Govekar

PROVIDER: PXD056828 | Pride | 2025-10-30

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	checksum.txt	Txt
	igg_ip-control_set1_1.1-ip-c_1.1.mzml	Mzml
	igg_ip-control_set1_1.1.group	Other
	igg_ip-control_set1_1.1.wiff	Wiff
	igg_ip-control_set1_1.1.wiff.scan	Wiff

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Project description:Sensitivity and throughput of transcriptomics and proteomomics technologies have advanced tremendously in recent years. With the use of deep sequencing of RNA samples (RNAseq) and mass spectrometry technology for protein identification and quantitation, it is now feasible to compare gene and protein expression on a massive scale and on any organism for which genomic data is available. Although these technologies are currently applied to many research questions in various model systems ranging from cell cultures to the entire organism level, there are few comparative studies of these technologies in the same system, let alone on the same samples. Here we present a comparison between gene and protein expression in zebrafish embryos, which is an upcoming model in disease studies. We compared Agilent custom made expression microarrays with Illumina deep sequencing for RNA analysis, showing as expected a high degree of correlation of expression of a common set of 15,927 genes. Gene expression was also found to correlate with the abundance of 1,017 quantified proteins, with several categories of genes as exceptions. These exceptions include ribosomal proteins, histones and vitellogenins, for which biological and technical explanations are discussed. By comparing state of the art transcriptomic and proteomic technologies on samples derived from the same group of organisms we have for the first time benchmarked the differences in these technologies with regard to sensitivity and bias towards detection of particular gene categories. Our datasets submitted to public repositories are a good starting point for researchers interested in disease progression in zebrafish at a stage of development highly suited for high throughput screening technologies. The transcriptomics data is deposited in NCBI GEO. Proteomics informatics: Spectra were identified by Mascot and validated by PeptideProphet and ProteinProphet in the Trans-Proteomic Pipeline (TPP). The main search parameters are: full tryptic specificity, precursor mass searched within -0.5 to +2.5 Da, product ions within [-0.5,0.5] Da and allowing for one missed cleavage. Carbamidomethylation was assumed as the only and fixed PTM.

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Immunoprecipitation followed by mass spectrometry (IP-MS)

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