title: Medsaic Human DotScan antibody microarray v3
status: Public on Feb 20 2013
contact.city: Boston
contact.first: Joshua
contact.middle: WK
contact.last: Ho
contact.email: jwho@rics.bwh.harvard.edu
contact.state: MA
contact.address: 41 Avenue Louis Pasteur
contact.department: Department of Medicine
contact.country: USA
contact.institute: Brigham and Women's Hospital
organism: Homo sapiens
manufacture_protocol: The use of CD antibody (DotScan) microarrays (Medsaic, veleigh, NSW, Australia) has been previously described. Briefly, they consist of grid pattern of antibody spots, each approximately 450 mm in diameter, robotically arrayed on a NC patch bound to a glass slide (FAST slides; supplied by Schleicher and Schuell, Keene, NH). The ability of an antibody spot to bind leukocytes depends on the number and availability of its corresponding antigens. Eighty-two mouse antihuman mAb spots were printed in duplicate on the slide, resulting in identical left and right panels of antibodies. Each spot contains a different CD antibody (the remaining spaces were reserved for testing new antibodies). Below the 82 spots is a smaller panel of monoclonal and polyclonal antimouse antibodies which serve as negative controls (not shown). Nonspecific binding to antibody spots was minimized by blocking slides with 5% w/v skim milk (Diploma; Bonlac Foods, Melbourne, Vic., Australia) in PBS overnight at 47C. After blocking, slides were washed twice in water, air-dried, and stored at 47C with desiccant. Processing of the microarrays and identification of spots has been described elsewhere.