|
| Status |
Public on Feb 02, 2014 |
| Title |
KPEpitopset1_spec_rep1 [4_1] |
| Sample type |
protein |
| |
|
| Source name |
proteins
|
| Organism |
Klebsiella pneumoniae |
| Characteristics |
strain: MGH 78578
|
| Extracted molecule |
protein |
| Extraction protocol |
N/A
|
| Label |
Chromeo-546
|
| Label protocol |
N/A
|
| |
|
| Hybridization protocol |
Slides were blocked using SuperBlock (Pierce) for 2 h at room temperature with mild shaking. Incubation with primary antibody (rabbit polyclonal to C.jejuni, E.coli or S.enterica) was performed overnight at 4°C with mild shaking in a humidity chamber. Secondary antibody (goat polyclonal to rabbit, Chromeo-546 conjugated) was incubated for 1.5 h with mild shaking at room temperature in the dark. After each incubation step, the slides were washed three times with PBSI (PBS + 0.05% IGEPAL). Slides were dried with nitrogen and scanned afterwards.
|
| Scan protocol |
Fluorescent array images were collected for Cy3 with a Genepix 4200A fluorescent scanner and a PMT gain of 400. Image intensity data were extracted and analyzed using Origin Pro 8G.
|
| Description |
raw data file: 4.txt
15mer oligopeptides with an overlap of 11 residues comprising the entire sequence of a set of proteins from K. pneumoniae MGH 78582
|
| Data processing |
From the raw data the corrected median fluorescent intensities (F532 median - B532) were used. Then, relative fluorescence intensities were calculated by subtracting the fluorescent intensities of the chamber incubated with secondary antibody from the other chambers. This was done for all the samples except the positive control Rabbit IgG, as the secondary antibody (Goat antibody to Rabbit IgG) is supposed to bind at this site. This acknowledges the presence of unspecific interaction of the secondary antibody to the samples.
|
| |
|
| Submission date |
Nov 19, 2013 |
| Last update date |
Feb 02, 2014 |
| Contact name |
Sebastian Hoppe |
| E-mail(s) |
sebastian.hoppe@izi-bb.fraunhofer.de
|
| Organization name |
Fraunhofer Institute for Cell Therapy and Immunology
|
| Department |
Bioanalytics and Biosensorics
|
| Lab |
Molecular Bioengineering
|
| Street address |
Am Mühlenberg 13
|
| City |
Potsdam |
| ZIP/Postal code |
14476 |
| Country |
Germany |
| |
|
| Platform ID |
GPL17960 |
| Series (1) |
| GSE52537 |
Epitope Mapping of potential novel immunogenic proteins of Klebsiella pneumoniae MGH 78578 |
|
