|
Status |
Public on Mar 01, 2014 |
Title |
Lu_24C3 |
Sample type |
RNA |
|
|
Source name |
Lung Control 24h Animal # 99
|
Organism |
Mus musculus |
Characteristics |
background strain: C3H/HeJ gender: Male tissue: Lung
|
Treatment protocol |
Animals were anesthetized and SEB toxin (treatment) or saline (control) was administered i.n. (5 μg SEB/ 50 μl) followed by and i.p. (2 μg SEB/200 μ-l) dose 2h later. PBMC, spleen, lung, liver, kidney, and heart were harvest for tRNA purification from SEB and saline control groups at 0, 2.75, 5 and 24 h after treatment .
|
Growth protocol |
Male C3H/HeJ, weighing ~ 20 g each (7-10 weeks old), were housed in conventional micro isolator cages. Food and water were freely available at all times.
|
Extracted molecule |
total RNA |
Extraction protocol |
PBMC were isolated by gradient (Nycoprep) from blood collected in EDTA-treated tubes and lysed in RLT buffer. Tissues were thinly sliced and preserved in RNAlater reagent at -80 C. tRNA was obtained using RNeasy kit (Qiagen) following manufacturer protocols. tRNA concentration was measure by nanodrop and tRNA quality was evaluated by Bioanalyzer.
|
Label |
Biotin
|
Label protocol |
Affymetrix GeneChip Expression 3'-Amplification One-Cycle Target Labeling and Control Kit was used for the synthesis of cDNA and cRNA and biotin-labeled cRNA. 2 μg of tRNA was utilized as starting material following manufacturer protocol.
|
|
|
Hybridization protocol |
20 μg of fragmented biotin-labeled cRNA was used in hybridization cocktails on 430 2.0 Mouse Genome Arrays (Affymetrix) following manufacturer protocols.
|
Scan protocol |
Affymetrix 430 2.0 array chips were scanned using the Affymetrix GeneChip Scanner 7G.
|
Description |
RLD_Nov08_Lung_24h_CTRL-3
|
Data processing |
Data were generated using the Mas 5.0 algorithm in the Affymetrix Expression Console software
|
|
|
Submission date |
Nov 18, 2013 |
Last update date |
Dec 12, 2023 |
Contact name |
Gabriela A Ferreyra |
E-mail(s) |
gferreyra@cc.nih.gov
|
Phone |
301-435-2317
|
Organization name |
Clinical Center
|
Department |
CCMD
|
Street address |
9000 Rockville Pike - Building 10 room 4D03
|
City |
Bethesda |
State/province |
MD |
ZIP/Postal code |
20892 |
Country |
USA |
|
|
Platform ID |
GPL1261 |
Series (1) |
GSE52474 |
Late Multiple Organ Surge in Interferon-regulated Target Genes Characterizes Staphylococcal Enterotoxin B Lethality |
|