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Experiment was performed on 293 cells, details are presented in publication: Adenoviral E4 gene stimulates secretion of PEDF that maintains long-term survival of human glomerulus -derived endothelial cells Marina Jerebtsova‡§*, Namita Kumari‖, Yuri Obuhkov ‖, and Sergei Nekhai‖†*
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Experiment was performed on 293 cells, details are presented in publication: Adenoviral E4 gene stimulates secretion of PEDF that maintains long-term survival of human glomerulus-derived endothelial cells Marina Jerebtsova‡§*, Namita Kumari‖, Yuri Obuhkov‖, and Sergei Nekhai‖†* (to be published in Molecular and Cellular Proteomics), please see more details in paper
Sample: 293 cells conditioned by E4 • Raw Excalibur spectra files represented by files MJ_1_9.raw,..MJ_1_16.raw • Proteome Discovere 1.2 SEQUEST search files represented by MJ_1_9.msf,.., MJ_1_16.msf • Control: 293 cells • Raw Excalibur spectra files represented by files SN_2_9.raw,..SN_1_18.raw • Proteome Discovere 1.2 SEQUEST search files represented by SN_2_9.msf,.., SN_2_18.msf • SEQUEST search was made on human.fasta database. • Each file corresponds to gel band and set of files represent two samples MJ and SN respectively
Procedure and tests • Workflow • HPLC on PicoFrit C18 column, gradient 60 min from 2% to 30% ACN • Nanospray 2 kV, flow rate 300 nL/min • LTQ Orbitrap MS. Data dependent scan FT MS + 3 FT MS/MS. • A blank water run was performed after each sample run • Each 24 hours setup performance was tested on Apomyoglobin sample, 12 fmol. Coverage was 39-79 % • Search is made by SEQUEST on human database