|
| Status |
Public on Aug 14, 2013 |
| Title |
RNA from Immunoprecipitation with IgG Antibody |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Proliferating C2C12
|
| Organism |
Mus musculus |
| Characteristics |
antibody: IgG
cell line: C2C12
|
| Treatment protocol |
No treatments
|
| Growth protocol |
Proliferating C2C12 muscle cells were grown in DMEM supplemented with 20% FBS and 1% Pen/Strep. At 80% confluency, cells were collected.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted with phenol/chloroform from proliferating C2C12 cell extracts that were subject to immunoprecipitation with an antibody that recognized either HuR or IgG. Total RNA was isolated from proliferating C2C12 using TRIzol reagent and used as reference sample.
|
| Label |
Hy3
|
| Label protocol |
The miRCURY™ LNA Array power labelling kit was used to label 600 - 1000ng of RNA isolated from immunoprecipitation with Hy3 and reference samples from total RNA with Hy5.
|
| |
|
| Channel 2 |
| Source name |
Common Reference Pool
|
| Organism |
Mus musculus |
| Characteristics |
cell line: C2C12
antibody: none
|
| Treatment protocol |
No treatments
|
| Growth protocol |
Proliferating C2C12 muscle cells were grown in DMEM supplemented with 20% FBS and 1% Pen/Strep. At 80% confluency, cells were collected.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted with phenol/chloroform from proliferating C2C12 cell extracts that were subject to immunoprecipitation with an antibody that recognized either HuR or IgG. Total RNA was isolated from proliferating C2C12 using TRIzol reagent and used as reference sample.
|
| Label |
Hy5
|
| Label protocol |
The miRCURY™ LNA Array power labelling kit was used to label 600 - 1000ng of RNA isolated from immunoprecipitation with Hy3 and reference samples from total RNA with Hy5.
|
| |
|
| |
| Hybridization protocol |
Hy3™-labeled immunoprecipitated RNA and Hy5™-labeled reference RNA were mixed pair-wise and hybridized to the miRCURY™ LNA array version 11.0 according to the miRCURY™ LNA array manual using a Tecan HS4800 hybridization station. Hybridized slides were stored in an ozone-free environment (ozone levels below 2.0 ppb)
|
| Scan protocol |
Scanned using the Agilent G2565BA Microarray Scanner System (Agilent Technoliges, Inc., USA) in an ozone-free environment
|
| Description |
RNA from immunoprecipitation using proliferating C2C12 extract and anti-IgG antibody
|
| Data processing |
Image analysis was carried out using the ImaGene 8.0 software (BioDiscovery, Inc.,USA). The quantified signals were background corrected (Normexp with offset value 10 according to the convolution method described by Ritchie ME, Bioinformatics (2007), Vol. 23, no. 20, pages 2700-2707. The signals were further normalized using the global Lowess (LOcally WEighted Scatterplot Smoothing) regression algorithm.
|
| |
|
| Submission date |
Aug 13, 2013 |
| Last update date |
Aug 14, 2013 |
| Contact name |
Imed Gallouzi |
| E-mail(s) |
imed.gallouzi@mcgill.ca
|
| Phone |
514-398-4537
|
| Organization name |
McGill University
|
| Department |
Biochemistry
|
| Lab |
915
|
| Street address |
3655 Promenade Sir William Osler
|
| City |
Montreal |
| State/province |
QC |
| ZIP/Postal code |
H3G1Y6 |
| Country |
Canada |
| |
|
| Platform ID |
GPL17568 |
| Series (2) |
| GSE49856 |
HuR and miR-1192 regulate myogenesis by modulating the translation of HMGB1 mRNA (mRNA) |
| GSE49857 |
HuR and miR-1192 regulate myogenesis by modulating the translation of HMGB1 mRNA |
|
