|
| Status |
Public on Dec 06, 2012 |
| Title |
Cultured GRK2-expressing HEK cell clone-rep2 |
| Sample type |
RNA |
| |
|
| Source name |
Cultured HEK cell clone with GRK2 expression
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: HEK cell clones
genotype/variation: GRK2
protocol: cultured in vitro
|
| Treatment protocol |
For the microarray study, cell pellets were isolated from three study groups: (i) NOD.Scid mouse-expanded HEK clones expressing either GRK2 or kinase-deficient GRK2-K220R (Scid), (ii) in vitro cultured HEK cell clones expressing GRK2 or GRK2-K220R (HEK), and (iii) in vitro re-cultured HEK cell clones after NOD.Scid mouse expansion (Ex-Scid).
|
| Growth protocol |
The study was performed with HEK cell clones stably expressing GRK2 or the kinase-deficient GRK2-K220R mutant. HEK cell clones were either expanded for 4 weeks in vivo in immunodeficient NOD.Scid mice (3 months of age), cultured in vitro, or re-cultured in vitro after expansion in NOD.Scid mice
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated from HEK cell clones with the RNeasy Mini kit according to the manufacturer`s instructions (Qiagen).
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA was prepared according to the protocol of the manufacturer (Affymetrix; GeneChip Expression Analysis Technical Manual).
|
| |
|
| Hybridization protocol |
For hybridization, 15 µg of fragmented cRNA were incubated with the chip (Affymetrix Human genome U133 Plus 2.0 Array) in 200 µl of the hybrization solution in a Hybridization Oven 640 (Affymetrix) for 16 h at 45 °C. GeneChips were then washed and stained using the Affymetrix Fluidics Station 450 according to the GeneChip Expression Analysis Technical Manual (Rev. 5).
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GeneChip Scanner 7G.
|
| Description |
HEK-GRK-2
Cultured GRK2-expressing HEK cell clone
Gene expression data of cultured GRK2-expressing HEK cell clone
|
| Data processing |
The signals were processed using Affymetrix GeneChip Operating Software (GCOS; v.1.4; Affymetrix). To compare samples and experiments, the trimmed mean signal of each array was scaled to a target intensity of 200.
|
| |
|
| Submission date |
Dec 06, 2012 |
| Last update date |
Dec 06, 2012 |
| Contact name |
Ursula Quitterer |
| E-mail(s) |
ursula.quitterer@pharma.ethz.ch
|
| Phone |
+41-446329801
|
| Organization name |
ETH Zurich
|
| Department |
Molecular Pharmacology
|
| Street address |
Winterthurerstrasse 190
|
| City |
Zurich |
| ZIP/Postal code |
CH-8057 |
| Country |
Switzerland |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE42771 |
Microarray gene expression profiling of kinase-dependent and kinase-independent effects of GRK2 |
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