|
| Status |
Public on May 01, 2012 |
| Title |
Parathyroid - Prolactin 3 h - Replicate 2 |
| Sample type |
RNA |
| |
|
| Source name |
Parathyroid_200 mg/L Prolactin_3h
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: Parthyroid Adenoma
|
| Treatment protocol |
Expression profiling was done in parathyroid adenomas subjected to prolactin treatment in culture. In addition, corresponding paraffin sections were obtained for verification of PRLr expression by immunohistochemistry. 200 mg/L prolactin (recombinant human prolactin, Cat. No. JM-4687-50, MBL Woburn, MA) was added to 1× 10^6 attached parathyroid tumour cells. Cells were harvested using RNAlater (QIAGEN) and homogenized with QIAshredder for RNA extraction after 3 h and 24 h in culture, respectively. Negative controls were collected in parallel with each case at the same time points. RNA was extracted using QIA Cube, and quality assessed with Bioanalyser and Nanodrop.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Parathyroid adenoma samples for culturing and functional studies were obtained at surgery. Up to one third of the tumour was collected from each sample, while the majority of the tissue was sent to routine histopathological diagnostics. Fresh tissue samples were quickly transferred in cold culturing media to the nearby laboratory, and then isolated and cultured using previously described methods. All experiments were performed within 72 hours after isolation.
|
| Label |
biotin
|
| Label protocol |
Total RNA was converted to sense strand cDNA using Ambion WT Expression kit. For fragmentation and labeling WT GeneChip WT target Labeling kit was used.
|
| |
|
| Hybridization protocol |
Hybridization to Affymetrix Human Gene 1.1 ST Array plates was performed using GeneTitan Hybridization, Wash, and Stain Kit for WT Array Plates.
|
| Scan protocol |
Array plates were scanned using Affymetrix GeneChip HT Array Plate Scanner according to standard protocols (Affymetrix Inc., Santa Clara, CA).
|
| Description |
Gene expression of parathyroid adenoma
|
| Data processing |
Preprocessing was performed in Affymetrix Expression Console (v. 1.1) using the following methods: Summarization: PLIER, Background Correction: PM-GCBG, Normalization: Global Median.
|
| |
|
| Submission date |
Sep 26, 2011 |
| Last update date |
May 01, 2012 |
| Contact name |
Felix Haglund |
| Organization name |
Karolinska Institutet
|
| Department |
MMK
|
| Lab |
C Larsson
|
| Street address |
Karolinska Universitetssjukhuset Solna, CMM L8:01
|
| City |
Stockholm |
| ZIP/Postal code |
17176 |
| Country |
Sweden |
| |
|
| Platform ID |
GPL11532 |
| Series (1) |
| GSE32387 |
Prolactin stimulation of parathyroid adenomas |
|
