1. How Do Antibody Arrays Work?

2. Antibodies Are Robotically Arrayed on a Slide Anti CD-AntibodiesAnti-CD Antibodies Nitrocellulose coat Glass slide Robot deposits 10 nL of antibody 400-450 µ spot Robot deposits 10 nL of antibody 400-450 µ spot

3. Leukocytes are Added to the Slide (usually PBMCs and added separately from PMNs) Anti CD Antibodies Different Leukocytes (e.g. T cells) Express Different CD Antigens

4. Leukocytes Bind to CD Antibodies (Depending on the match between Ab and Epitope) Anti CD Antibodies Leukocytes with non- matching epitopes do not bind Leukocytes with matching antigens are captured

5. Leukocytes That Are Not Captured Are Washed Away Anti CD-Y AntibodiesAnti CD-X Antibodies About 1000 cells are captured on any one antibody spot

6. CD Antibody Microarrays display a pattern of cell capture that distinguishes between healthy and inflammatory conditions LymphocytesGranulocytes CD antibodies in the array