|
Status |
Public on Feb 21, 2013 |
Title |
120674_Other_Autoimmune |
Sample type |
protein |
|
|
Source name |
Peripheral Blood Mononuclear Cells
|
Organism |
Homo sapiens |
Characteristics |
cell type: PBMC diagnosis: Other_Autoimmune
|
Extracted molecule |
protein |
Extraction protocol |
Anticoagulated whole blood was collected from each participant in EDTA and maintained at room temperature until use within 24 hours. Leukocytes were isolated from peripheral blood using Histopaque (Sigma-Aldrich), washed in PBS, resuspended in PBS containing 1 mM EDTA to a density of 107 cells/ml, and incubated on the cell array for 30 min at room temperature (100 ml of suspension/slide), and unbound cells were then gently washed off with PBS. Arrays were then fixed for at least 1 hour in PBS containing 1% (v/v) formaldehyde (Sigma-Aldrich) and washed in PBS, scanned and the light scattering by the attached cells recorded in an image file.
|
Label |
none
|
Label protocol |
No labeling
|
|
|
Hybridization protocol |
Briefly, duplicate patterns of anti-CD antibody spots were robotically arrayed onto nitrocellulose membrane coated glass slides (Schleicher and Schuell, Keene, NH), and blocked with skim milk (5% in PBS).
|
Scan protocol |
Cells captured on antibody dots were imaged using a DotScan™ slide reader (Medsaic, NSW, Australia). The number of immobilized cells was proportional to the light scattered at each antibody spot. Binding of PBMCs to each antibody dot was monitored by light scatter and averaged between the duplicate spots on each slide.
|
Description |
This array survey leukocyte surface CD markers
|
Data processing |
Data are scale-normalized such that the spot values of each array ranges between 0 and 10 with respect to the spot with the maximum binding density. Only spots with signal-to-noise ratio greater than 1.2 and upper average expression greater than 1 are included for further analysis, and are reported here.
|
|
|
Submission date |
Feb 14, 2011 |
Last update date |
Feb 21, 2013 |
Contact name |
Joshua WK Ho |
E-mail(s) |
jwho@rics.bwh.harvard.edu
|
Organization name |
Brigham and Women's Hospital
|
Department |
Department of Medicine
|
Lab |
Division of Genetics
|
Street address |
41 Avenue Louis Pasteur
|
City |
Boston |
State/province |
MA |
ZIP/Postal code |
02115 |
Country |
USA |
|
|
Platform ID |
GPL13165 |
Series (1) |
GSE27293 |
An antibody leukocyte capture microarray in diagnosis of Systemic Lupus Erythematosus |
|