|
| Status |
Public on Dec 14, 2010 |
| Title |
Jurkat_T_Cells_stimulated_4H_PKC-theta_rep1_slide2of2 |
| Sample type |
genomic |
| |
|
| Channel 1 |
| Source name |
Total IP 4H
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: Jurkat T Cells - stimulated
age: 4 hours
|
| Treatment protocol |
ChIP assays were performed following the protocol supplied by Upstate Biotechnology, as previously detailed (MCB 2009).
|
| Growth protocol |
Human Jurkat T cells were cultured in RPMI 1640 medium (Gibco) supplemented with 10% FCS, 2mM L-glutamine, 9 mM HEPES and antibiotics. Jurkat T cells were maintained in suspension at a density of 5 x 10^5 cells/ml. Cells were stimulated with 20 ng/ml of phorbol 12-myristate 13-acetate (P) (Boehringer Mannheim) and 1 µM Ca 2+ ionophore A23187 (I) (Sigma-Aldrich) for the times indicated.
|
| Extracted molecule |
genomic DNA |
| Extraction protocol |
Pooled ChIP samples were subsequently amplified based on one round of the whole genome amplification method using the WGA2 kit (Sigma-Aldrich), as described previously (O'Geen, Hollenhorst, Dindot)
|
| Label |
Cy3
|
| Label protocol |
Labelling, hybridization and scanning were performed as described in the Mammalian ChIP-on-chip protocol (version 9.1; Agilent Technologies)
|
| |
|
| Channel 2 |
| Source name |
PKC-theta 4H
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: Jurkat T Cells - stimulated
age: 4 hours
antibody: anti-PKC-theta
sample type: DNA pooled from five independent ChIP assay experiments
|
| Treatment protocol |
ChIP assays were performed following the protocol supplied by Upstate Biotechnology, as previously detailed (MCB 2009).
|
| Growth protocol |
Human Jurkat T cells were cultured in RPMI 1640 medium (Gibco) supplemented with 10% FCS, 2mM L-glutamine, 9 mM HEPES and antibiotics. Jurkat T cells were maintained in suspension at a density of 5 x 10^5 cells/ml. Cells were stimulated with 20 ng/ml of phorbol 12-myristate 13-acetate (P) (Boehringer Mannheim) and 1 µM Ca 2+ ionophore A23187 (I) (Sigma-Aldrich) for the times indicated.
|
| Extracted molecule |
genomic DNA |
| Extraction protocol |
Pooled ChIP samples were subsequently amplified based on one round of the whole genome amplification method using the WGA2 kit (Sigma-Aldrich), as described previously (O'Geen, Hollenhorst, Dindot)
|
| Label |
cy5
|
| Label protocol |
Labelling, hybridization and scanning were performed as described in the Mammalian ChIP-on-chip protocol (version 9.1; Agilent Technologies)
|
| |
|
| |
| Hybridization protocol |
Labelling, hybridization and scanning were performed as described in the Mammalian ChIP-on-chip protocol (version 9.1; Agilent Technologies)
|
| Scan protocol |
Scanned on Agilent G2565BA Scanner
|
| Description |
Biological replicate 1 of 2 on the second of the two slides that comprise a single 2-colour ChIP-chip experiment
|
| Data processing |
Scanned images were quantified with Agilent Feature Extraction software under the default chip-chip protocol (version 9.5.3).
|
| |
|
| Submission date |
Dec 13, 2010 |
| Last update date |
Dec 14, 2010 |
| Contact name |
Russell Leigh McInnes |
| Organization name |
Agilent Technologies
|
| Department |
LSCA
|
| Street address |
347 Burwood Hwy
|
| City |
Forest Hill |
| State/province |
Vic |
| ZIP/Postal code |
3131 |
| Country |
Australia |
| |
|
| Platform ID |
GPL4125 |
| Series (1) |
| GSE26035 |
Chromatin-associated protein kinase C-0 regulates an inducible gene expression program and microRNAs in human T lymphocytes |
|
