Kinome analysis of human CD151+ and CD151- T cells in the absence or presence of IL-2 stimulation

ABSTRACT: Primary human T cells from healthy donors were sorted using a FACSAria II to obtain CD3+CD4+CD151+ and CD3+CD4+CD151- populations. To perform kinome analysis using Kinex™ microarrays, 50 µg of lysate protein from each sample were labeled with a proprietary fluorescent dye according to the manufacturers instructions (Kinexus, Canada). The utilized KAM-850 chips were spotted in duplicates with over 850 antibodies: 510 pan-specific antibodies used in the chip allows for the detection of 189 protein kinases, 31 protein phosphatases and 142 regulatory subunits of these enzymes and other cell signaling proteins. 340 phospho-specific antibodies tracked the unique phosphorylation of 128 sites in protein kinases, 4 sites in protein phosphatases and 155 sites in other cell signaling proteins. The background-corrected raw intensity data were logarithmically transformed with base 2. Z scores were calculated by subtracting the overall average intensity of all spots within a sample from the raw intensity for each spot, and Z score ratios were calculated by dividing Z scores by the standard deviations (SD) of all of the measured intensities within each sample. To minimize unspecific background signals, lysates from cells from 5 different donors were sorted and the corresponding populations were pooled to provide one sample prior to loading on the Kinexus antibody microarrays. 16JN20-KAM880 Kit-K012003211-19123.txt contains the original data for the CD4+CD151- T cell population at baseline. 16JN20-KAM880 Kit-K012003211-19124.txt contains the original data for the CD4+CD151- T cell population following stimulation with 30 U/ml IL-2 for 24 hours. 16JN21-KAM880 Kit-K012003229-19125.txt contains the original data for the CD4+CD151+ T cell population at baseline. 16JN21-KAM880 Kit-K012003229-19126.txt ontains the original data for the CD4+CD151+ T cell population following stimulation with 30 U/ml IL-2 for 24 hours.

ORGANISM(S): Homo sapiens

SUBMITTER: Olaf Kutsch 

PROVIDER: E-MTAB-5977 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress