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Status |
Public on Feb 01, 2022 |
Title |
Control 1, WB α-casein stimulation |
Sample type |
RNA |
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Source name |
Control 1, WB α-casein stimulation
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Organism |
Homo sapiens |
Characteristics |
tissue: whole blood gender: male age(day): 790
|
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.
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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.
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Scan protocol |
Fluorescence images of the hybridized arrays were generated using the Agilent Scanner (Scan control software ver. 8)
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Description |
Gene expression, whole blood sample from control1 was stimulated with α-casein for 24 hours
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Data processing |
The intensities were extracted with Agilent Feature Extraction software ver. 10.5.1.1
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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. |
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