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Sample GSM499997 Query DataSets for GSM499997
Status Public on Jan 03, 2012
Title Serum sample 6
Sample type protein
 
Source name Human sera
Organism Homo sapiens
Characteristics fluid: Human serum
gender: F
clinical status: asthmatic
eczema: undefined diagnosis
conjunctivitis: undefined diagnosis
rhinitis: undefined diagnosis
Extracted molecule protein
Extraction protocol At the recruitment sessions, each subject was interviewed, disease status was ascertained by physical examination, permission was asked to access personal health records, and blood samples were collected
Label Alexa 555
Label protocol TSA™ Kit #42 *with HRP—streptavidin and Alexa Fluor® 555 tyramide* *50-150 slides* (Invitrogen)
 
Hybridization protocol Printed slides were blocked with PBS containing 2% BSA for 1 h at room temperature. Slides were then incubated with serum samples (100 µl) for 60 minutes at 37°C. To reveal bound IgE, the slides were incubated with a secondary mouse monoclonal antibody directed against human IgE (0.14 µg/ml - 100µl) for 45 minutes at 37°C, followed by an incubation with anti-mouse IgG HRP conjugated antibody (1.6 µg/mL - 100µl) for 45 minutes at 37°C and finally incubated with tyramide-Alexa 555 (TSA™ Kit #42 *with HRP—streptavidin and Alexa Fluor® 555 tyramide* *50-150 slides*) (Invitrogen) diluted 1:200 (100µl), for 15 minutes at 37°C. Slides were dried at 37°C before measuring the fluorescence signal.
Scan protocol The processed slides were scanned using a fluorescence-detecting scanner ScanArray™ Gx and the images were generated with the ScanArray™ software provided by Perkin Elmer Life Sciences Inc. All the slides were scanned under identical settings: 90 % laser power and 60 % photomultiplier gain.
Description no additional information
Data processing Quantification of bound IgE: The fluorescence signal was acquired using ProScanArray Express™ version 3.0 software. PMC reading values of individual spots were corrected against the internal negative control to identify signals above background. Duplicate measurements of individual allergens were utilized. The signal collected from the allergens was interpolated with an external calibration curve to obtain the IU/ml value (see supplementary file GSE20020_IU_ml values.txt on the GSE20020 record), and translated into a Class Score by plotting the data in a standard reactivity scale (see Sample data tables). Class Score values: CLASS 0 (less than 0.35 IU/ml); CLASS 1 (0.35-0.7 IU/ml); CLASS 2 (0.71-3.5 IU/ml); CLASS 3 (3.51-17.5 IU/ml); CLASS 4 (17.51-50 IU/ml); CLASS 5 (50.01-100 IU/ml).
 
Submission date Jan 24, 2010
Last update date Jan 03, 2012
Contact name tania dottorini
Organization name University of Perugia
Department Dept. Exp.Medicine and Bichem. Sci.
Street address via del Giochetto
City Perugia
ZIP/Postal code 06126
Country Italy
 
Platform ID GPL9968
Series (1)
GSE20020 IgE reactivity profiling in an asthma affected cohort

Data table header descriptions
ID_REF
VALUE Standard reactivity class score scale, as outlined in 'Data processing' field.

Data table
ID_REF VALUE
C1 1
C2 0
C214 1
D01 1
D02 0
D03 0
D70 0
D71 0
D72 0
D73 0
E01 0
E02 0
E03 0
E06 0
E78 0
E81 0
E82 0
F01 0
F02 2
F03 1

Total number of rows: 103

Table truncated, full table size <1 Kbytes.




Supplementary file Size Download File type/resource
GSM499997.xls.gz 32.2 Kb (ftp)(http) XLS
Processed data included within Sample table

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