|
| Status |
Public on Oct 01, 2013 |
| Title |
immunoscreening S. Enteritidis 2 (416927) |
| Sample type |
protein |
| |
|
| Source name |
crude lysate Acella E.coli + recombinantly expressed proteins
|
| Organism |
Salmonella enterica subsp. enterica serovar Enteritidis |
| Characteristics |
strain: 125109
|
| Extracted molecule |
protein |
| Extraction protocol |
Fusion proteins were extracted from cultures using EasyLyse Bacterial protein extraction kit (Epicentre)
|
| Label |
Chromeo-546
|
| Label protocol |
n/a
|
| |
|
| Hybridization protocol |
Fusion proteins were spotted and coupled to the microarray surface for one hour at room temperature with 70% humidity. Slides were washed three times with PBST (PBS + 0.01% Tween20). Incubation with primary antibody (rabbit polyclonal to S. enterica) and secondary antibody (goat polyclonal to rabbit, Chromeo-546 conjugated) were performed for 1.5 h each, with washing (three times) with PBST after each incubation step. Slides were dried with nitrogen and scanned afterwards.
|
| Scan protocol |
Fluorescent array images were collected for Cy3 with a Genepix 4200A fluorescent scanner. Image intensity data were extracted and analyzed using Origin Pro 8G.
|
| Description |
HaloTag-fusion proteins derived from Campylobacter jejuni cDNA and expressed in Acella E.coli overexpression strains.
|
| Data processing |
From the raw data the corrected median fluorescent intensities (F532 median - B532) were used. Then, relative fluorescence intensities were calculated by subtraction the fluorescent intensities of the top compartment by the ones from the bottom compartment. This acknowledges the presence of unspecific interaction of the secondary antibody to the samples. Next, the contrast method was applied by calculating the contrast for each spot of the top compartment. C = [rfi(sample) - median rfi(negative reference)]/[rfi(sample) + median rfi(negative reference)]
|
| |
|
| Submission date |
Aug 06, 2013 |
| Last update date |
Oct 01, 2013 |
| 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 |
GPL17538 |
| Series (1) |
| GSE49605 |
Screening for novel immungenic proteins of Salmonella Enteritidis 125109 |
|
