|
| Status |
Public on Aug 09, 2019 |
| Title |
Mettl14 knock-out1 |
| Sample type |
protein |
| |
|
| Source name |
KO_islets
|
| Organism |
Mus musculus |
| Characteristics |
strain background: C57BL/6N
genotype/variation: KO; Mettl14 knock-out
tissue: islets
batch: A
|
| Extracted molecule |
protein |
| Extraction protocol |
Total proteins were harvested from islets lysates using M-PER protein extraction reagent (Thermo Fisher, USA) supplemented with proteinase and phosphatase inhibitors (Sigma, USA) according to standard protocol.
|
| Label |
biotin
|
| Label protocol |
Proteins were covalently labeled with biotin. Free biotin molecules are then removed at the completion of labeling reactions by gel filtration.
|
| |
|
| Hybridization protocol |
After blocking non-specific binding sites on the array, an incubation chamber is mounted onto the microarray to permit the loading of 2 samples side by side on the same chip and prevent mixing of the samples. Following sample incubation, unbound proteins are washed away and the array is then probed with anti-biotin antibody that is labelled with a proprietary fluorescent dye combination.
|
| Scan protocol |
Each array produces a pair of 16-bit images, which are captured with a Perkin-Elmer ScanArray Reader laser array scanner (Waltham, MA).
|
| Description |
The antibodies used on the Kinex™ KAM-1325 antibody microarrays consist of polyclonal and monoclonal antibodies that are specific for proteins or phosphosites.
|
| Data processing |
Signal quantification is performed with ImaGene 9.0 from BioDiscovery (El Segundo, CA) with predetermined settings for spot segmentation and background correction. The background-corrected raw intensity data are logarithmically transformed with base 2.
|
| |
|
| Submission date |
Jun 03, 2019 |
| Last update date |
Aug 09, 2019 |
| Contact name |
Hui Pan |
| E-mail(s) |
Hui.Pan@joslin.harvard.edu
|
| Organization name |
Joslin Diabetes Center
|
| Department |
Bioinformatics and Biostatistics Core
|
| Street address |
1 Joslin Place
|
| City |
Boston |
| ZIP/Postal code |
02215 |
| Country |
USA |
| |
|
| Platform ID |
GPL26730 |
| Series (1) |
| GSE132111 |
Phospho-antibody microarray analyses for islets of control and Mettl14 knock-out mice |
|
