|
| Status |
Public on Dec 01, 2008 |
| Title |
Patient_#4248_C3 |
| Sample type |
protein |
| |
|
| Source name |
serum
|
| Organism |
Homo sapiens |
| Characteristics |
anti-dsDNA:negative
SEX: Female
|
| Growth protocol |
Serum samples were stored at -70C
|
| Extracted molecule |
protein |
| Extraction protocol |
Serum sample was centrifuged for 6min at 15000g for removing particles
|
| Label |
Cy5
|
| Label protocol |
Labeling with fluorescent antibodies was carried out at room temperature for 30 min in 5% BSA and 0.05% Tween 20 containing PBS.
|
| |
|
| Hybridization protocol |
Dried arrays were rinsed in PBS for 15 minutes before use, then incubated with 5 times diluted serum at 37°C for 1 hour. Sera were diluted in 5% BSA, 0.05% Tween 20 Ca and Mg supplemented Veronal buffer. Serum treated slides were washed with 0.05% Tween 20 containing PBS, then incubated in 1:5000 diluted Cy5-conjugated F(ab')2 fragment goat anti-human IgM (Jackson ImmunoResearch). Following washing in 0.05% Tween 20 containing PBS, arrays were dried and scanned.
|
| Scan protocol |
Arrays were scanned on Typhoon Trio+ imager (Amersham Bioscience) (Excitation:633nm ; Emission: 670 BP30 , PMT:410 Mode:Fluorescence, Sensitivity:Normal, FocalPlane=+3 mm). Data were analyzed with ImageQuantTL (Amersham Bioscience) software.
|
| Description |
C3-4248
|
| Data processing |
Background derived from marked spots, was substracted from signal. For interassay comparison, data were normalized to the mean of two dilutions of printed anti-human IgG+IgM control material (ID_REF: #539, #571, #603, #355, #387, #419). Only data derived from slides with normalization factor between 0.4 and 2.5 were considered suitable for analysis. Based on the distribution of data normalized fluorescence intensity signals smaller than 1000 for IgM were rounded up to these values.
|
| |
|
| Submission date |
Sep 26, 2008 |
| Last update date |
Nov 13, 2008 |
| Contact name |
József Prechl |
| E-mail(s) |
jprechl@iif.hu
|
| Phone |
+3613812175
|
| Fax |
+3613812176
|
| URL |
http://www.abc-arrays.com
|
| Organization name |
ELTE-MTA
|
| Department |
Immunology Research Group
|
| Lab |
Immunoproteomics
|
| Street address |
Pázmány Péter s. 1/C
|
| City |
Budapest |
| ZIP/Postal code |
1117 |
| Country |
Hungary |
| |
|
| Platform ID |
GPL7390 |
| Series (1) |
| GSE12943 |
Profiling antibody function on antigen arrays |
|
