|
| Status |
Public on Feb 28, 2015 |
| Title |
pooled sera from six 8-10 month old BXD2 mice |
| Sample type |
other |
| |
|
| Source name |
BXD2 Mice Sera
|
| Organism |
Mus musculus |
| Characteristics |
strain: BXD2
disease model: Lupus
age: 8-10 mo
sample type: serum
|
| Extracted molecule |
other |
| Extraction protocol |
Sera were obtained from BXD2 mice by retro-orbital eye bleeding. After separation from blood, sera were stored at -80°C.
|
| Label |
Red DyLight680
|
| Label protocol |
The secondary antibodies used for all arrays were from Rockland Immunochemicals, Gilbertsville, PA. They were conjugated to the DyLight dyes (DyLight680 or DyLight800) by the company.
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| |
|
| Hybridization protocol |
Peptide arrays were first blocked 60 min in Blocking Buffer for Near Infra Red Fluorescent Western blotting (Rockland Immunochemicals, Inc., Gilbertsville, PA). The microarray was washed twice in PBS, pH 7.4 with 0.05% Tween 20 (PBS-T), and incubated for an additional 30 min in washing buffer. The array was then incubated overnight at 4°C with mouse sera diluted 1:1000 for anti-mouse IgG (H+L) analysis (Rockland Immunochemicals, Inc., Gilbertsville, PA). After multiple washes in washing buffer, the microarrays were incubated for 30 min with the secondary antibody in PBS-T at room temperature. After two additional washes in washing buffer, the microarrays were rinsed with ultrapure water and dried in a stream of air.
|
| Scan protocol |
Fluorescence intensities were acquired on an LI-COR Odyssey Imager (Lincoln, NE) at scanning intensity of 7, 0.8 - 1.0 mm offset, at a spatial resolution of 21 µm.
|
| Description |
Sample name: BXD2 Av. Foreground Median
|
| Data processing |
Quantification of spot intensities and peptide annotation were based on the 16-bit grey scale tiff file that exhibits a higher dynamic range than the 24-bit colorized tiff file. PepSlide® Analyzer software was used to analyze the data. A software algorithm breaks down fluorescence intensities of each spot into raw, foreground and background signal, and calculates the standard deviation of foreground median foreground intensities.
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| |
|
| Submission date |
Jan 26, 2015 |
| Last update date |
Feb 28, 2015 |
| Contact name |
Jennie Hamilton |
| Organization name |
University of Alabama Birmingham
|
| Department |
Department of Medicine, Division of Clinical Immunology and Rheumatology
|
| Street address |
1825 University Blvd.
|
| City |
Birmingham |
| State/province |
AL |
| ZIP/Postal code |
35294 |
| Country |
USA |
| |
|
| Platform ID |
GPL19700 |
| Series (2) |
| GSE65276 |
Autoimmune peptide antigen reactivity in BXD2 mice with spontaneous systemic autoimmune disease |
| GSE65290 |
General Approach for Tetramer Based Identification of Autoantigen Reactive B Cells: Characterization of La and snRNP Reactive B Cells in Autoimmune BXD2 Mice |
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