|
| Status |
Public on Aug 28, 2020 |
| Title |
ADO.021-PI |
| Sample type |
protein |
| |
|
| Source name |
vaccinated_Adolescent_serum-PI
|
| Organism |
Homo sapiens |
| Characteristics |
subject group: Adolescent
tissue: blood
tissue subtype: serum-PI
|
| Extracted molecule |
protein |
| Extraction protocol |
Human sera collected from different clinical trials
|
| Label |
AlexaFluor®647-labelled anti-human secondary antibody
|
| Label protocol |
The secondary antibody used for arrays was from Jackson Immunoresearch, (872 W Baltimore Pike, West Grove, PA 19390 USA). It was conjugated to the Alexafluor dye (Alexa 647) by the company.
|
| |
|
| Hybridization protocol |
Protein arrays were first blocked 60 min in Blocking Buffer (Block it, Arrayit Sunnyvale, CA 94085, USA). The array was then incubated 1h a room temperature with human sera diluted 1:500 for anti-human IgG-specific analysis. After washing with 0.1% Tween 20 in PBS buffer (PBST), arrays were incubated with an AlexaFluor®647-labelled anti-human IgG, fab specific secondary antibody (1:800 – Jackson Immunoresearch) at RT for 1 h to detect interactions. Slides were washed again as before, then rinsed in sterile water and dried.
|
| Scan protocol |
Fluorescence signals were detected by using a Powerscanner (Tecan Trading AG, Switzerland) and the 16-bit images were generated with Powerscanner software at 10 µm per pixel resolution.
|
| Description |
NCT00661713
FS00007597 Area1-ADO.021-PI
|
| Data processing |
Images were processed using ImaGene 9.0 software (Biodiscovery Inc, CA, USA). Elaboration and analysis of image raw fluorescence Intensity (FI) data was performed using in-house developed software and R scripts. For each sample, the mean fluorescence intensity (MFI) of replicated spots was determined, after subtraction of the background value surrounding each spot.
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| |
|
| Submission date |
Jun 18, 2020 |
| Last update date |
Aug 28, 2020 |
| Contact name |
Erika Bartolini |
| Organization name |
GSK Vaccines
|
| Department |
AIMB
|
| Lab |
Molecular Biology
|
| Street address |
Via Fiorentina 1
|
| City |
Siena |
| State/province |
Italy |
| ZIP/Postal code |
53100 |
| Country |
Italy |
| |
|
| Platform ID |
GPL28712 |
| Series (1) |
| GSE152785 |
Immunological fingerprint of 4CMenB recombinant antigens via protein microarray |
|
