|
Status |
Public on Dec 05, 2008 |
Title |
ACHN |
Sample type |
protein |
|
|
Source name |
renal
|
Organism |
Homo sapiens |
Characteristics |
renal
|
Growth protocol |
Cells from each cell line were grown on a 100mm dish to about 80% confluency.
|
Extracted molecule |
protein |
Extraction protocol |
Media were removed and ice-cold 1x cell lysis buffer were added (2ml per 100mm dish); lysates were collected by scrapping and then incubated on ice for 30min; after centrifuging at 13000rpm on a desktop centrifuge for 10min; supernatant was collected; the
|
Label |
biotinylated phospho-tyrosine antibodies
|
Label protocol |
According to manufacturer's protocol
|
|
|
Hybridization protocol |
antibody coupled beads were incubated with total protein lysates to capture targeted analytes; the beads were then labelled with biotinylated phospho-tyrosine antibodies and streptavidin, R-phycoerythrin conjugates
|
Scan protocol |
The data were collected with a Luminex 100 instrument
|
Description |
antibodies against the analytes were coupled to beads of corresponding color; total mouse, rat or rabbit IgG were used as negative controls; a phospho-tyrosine peptide was coupled to beads as a positive control raw data: D1 of Output 050307-3.csv
|
Data processing |
The background readings for each capture antibody were obtained using microspheres incubated with 1x cell lysis buffer (Cell Signaling Technology). Values were considered positive if they were 3-fold over the background and represented by log2-transformation of the folds over background. Negative values were threshold to -5.
|
|
|
Submission date |
Dec 03, 2008 |
Last update date |
Dec 04, 2008 |
Contact name |
Jinyan Du |
E-mail(s) |
jinyandu@broad.mit.edu
|
Phone |
617-324-4826
|
Organization name |
The Broad Institute
|
Department |
Cancer Program
|
Lab |
4175-NN
|
Street address |
7 Cambridge Center
|
City |
Cambridge |
State/province |
MA |
ZIP/Postal code |
02142 |
Country |
USA |
|
|
Platform ID |
GPL7696 |
Series (1) |
GSE13808 |
Bead-based kinase phosphorylation profiling identifies SRC as a therapeutic target in glioblastoma |
|