|
| Status |
Public on Nov 14, 2014 |
| Title |
A/Anhui/1/2013 |
| Sample type |
protein |
| |
|
| Channel 1 |
| Source name |
recombinant H7 HA from baculovirus
|
| Organism |
Influenza A virus |
| Characteristics |
strain: A/Anhui/1/2013
protein: A recombinant form of the H7 hemagglutinin (HA) protein from influenza virus
labeling: biotin then streptavidin-Cy5
|
| Extracted molecule |
protein |
| Extraction protocol |
recombinant HA from baculovirus
|
| Label |
Cy5
|
| Label protocol |
each recombinant HA was biotinylated overnight prior to incubation on the array
|
| |
|
| Channel 2 |
| Source name |
anti-human IgG Fcγ fragment specific
|
| Organism |
Capra hircus |
| Characteristics |
to normalize variation in antibody spotting: anti-human IgG antibody Fcγ fragment specific
|
| Extracted molecule |
protein |
| Extraction protocol |
anti-human IgG Fc fragment specific generated in goat
|
| Label |
Cy3
|
| Label protocol |
each recombinant HA was biotinylated overnight prior to incubation on the array
|
| |
|
| |
| Hybridization protocol |
The array was blocked for 1 hour. Then the anti-human IgG-Fc Cy3 was applied on the array for 1hour then washed. 40µg/ml of each biotinylated HA was applied to the array for 1hour then washed and finally the streptavidin-Cy5 was applied and then washed.
|
| Scan protocol |
scanned using GenePix 4000B
|
| Description |
raw data file: Wilson_11.12.14_array2.xlsx (Block 16)
Channel 1: recombinant HA biotinylated
Reactivity of the antibodies spotted on the array with the HA A/Anhui. Normalization with the anti-human IgG atibody Fcγ fragment specific.
For the HAs, A/Uruguay, A/Wisconsin, A/Perth, A/Victoria, A/Canada and A/Netherlands have been obtained by BEI ressources. From their website: The H3 hemagglutinin (HA) protein from influenza virus A/Wisconsin/67/2005 (H3N2) is a full-length glycosylated recombinant protein that was produced in Sf9 insect cells using a baculovirus expression vector system. The recombinant H3 HA protein was purified under conditions that preserve its biological activity and tertiary structure. For the HAs A/Anhui et A/Shanghai, they are home made but the same protocol was used.
|
| Data processing |
software GenePix Pro 6.0. Ratio of fluorescence (Cy5/Cy3) was used as the anti-human IgG Fc was used to normalize variation in antibody spotting.
|
| |
|
| Submission date |
Nov 13, 2014 |
| Last update date |
Nov 14, 2014 |
| Contact name |
Patrick C Wilson |
| E-mail(s) |
wilsonp@uchicago.edu
|
| Phone |
17737029009
|
| Organization name |
University of Chicago
|
| Department |
Medicine, Rheumatology
|
| Street address |
924 E 57th street, Knapp building
|
| City |
CHICAGO |
| State/province |
Illinois |
| ZIP/Postal code |
60637 |
| Country |
USA |
| |
|
| Platform ID |
GPL19411 |
| Series (1) |
| GSE63249 |
Identification of H3N2 specific antibodies cross-reacting with H7 strains |
|
