|
| Status |
Public on Feb 01, 2022 |
| Title |
Patient 3, WB non stimulation |
| Sample type |
RNA |
| |
|
| Source name |
Patient 3, WB non stimulation
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: whole blood
gender: female
age(day): 1215
|
| Treatment protocol |
The diluted blood was divided into each tube by 800 μL and it was incubated with α-casein (100 μg/mL; Sigma-Aldrich) or Pmix (100 μg/mL), which is a mixture of four milk protein components (α-casein, κ-casein, α-lactalbumin, or β-lactoglobulin; 100 µg/mL each; Sigma-Aldrich) at 37°C for 24 h
|
| Growth protocol |
Heparinized whole blood collected from patients with non-IgE GI-FAs and individuals who did not have non-IgE-GI-FAs or other immune diseases was diluted 4-fold with RPMI-1640 medium (Sigma-Aldrich, St. Louis, MO, USA).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNAs were extracted from the blood sample, which unstimulated or stimulated with milk protein by using Nucleo Spin® RNA Blood (MACHEREY-NAGEL GmbH & Co. KG, Düren Germany).
|
| Label |
Cy3
|
| Label protocol |
One-color spike-mix (Agilent Technology) was added to the total RNA prior to the labeling reaction. Labeling was performed using a Low lnput Quick Amp Labeling Kit (Agilent Technology) in the presence of cyanine-3 (Cy3)-CTP.
|
| |
|
| Hybridization protocol |
600 ug of Cy3-labelled cRNA was fragmented at 60°C for 30 minutes in a reaction volume of 25 μl containing 25x fragmentation buffer and 10x blocking agent following the manufacturers instructions. On completion of the fragmentation reaction, 25μl of 2x GE hybridization buffer HI-RPM was added to the fragmentation mixture and hybridized to SurePrint G3 Human GE Microarray 8 × 60 k ver. 2.0 (Design ID: 039494, Agilent Technologies) for 17 hours at 65°C.
|
| Scan protocol |
Fluorescence images of the hybridized arrays were generated using the Agilent Scanner (Scan control software ver. 8)
|
| Description |
Gene expression, whole blood sample from patient3 was not stimulated for 24 hours
|
| Data processing |
The intensities were extracted with Agilent Feature Extraction software ver. 10.5.1.1
|
| |
|
| Submission date |
Apr 07, 2021 |
| Last update date |
Feb 01, 2022 |
| Contact name |
Yag Hisako |
| E-mail(s) |
hisakoy2004@yahoo.co.jp
|
| Organization name |
Department of Pediatrics, Gunma University Graduate School of Medicine
|
| Street address |
3-39-22 Showa-machi
|
| City |
Maebashi |
| ZIP/Postal code |
371-8511 |
| Country |
Japan |
| |
|
| Platform ID |
GPL13607 |
| Series (1) |
| GSE171660 |
Novel Peripheral Blood Allergen Stimulation Test for Non-IgE-mediated Gastrointestinal Food Allergies in Neonates and Infants. |
|
