1. Peptide Microarray Service Binding buffer: 1X PBS (pH 7.0). Binding conditions: 1:100 diluted mouse serum samples in binding buffer, 4 ºC, 16 hours. Detection: 10 ng/mL anti-mouse IgG Cy5 conjugate in 1X PBS buffer, 4 ºC, 2 hours. Included are images after blocking and serum binding for each chip.

2. 01_120136-1203-block-700.tif12/03/2012 by AH Chip Pfizer_120136 To block chip surface: 1) Block with SuperBlock, 0.05% Tween-20, pH 7.0, 4 ºC overnight. 2) Wash with 1 mL of 1×PBS, pH7.0. 3) Image scanned at 635 nm and PMT 700. A B C D E F

3. 02_120136A-1206-1%Normal-700.tif, 02_120136B-1212-1%AR172-1-700.tif, 02_120136C-1212-1%AR172-3-700.tif 02_120136D-1214-1%AR188-2-700.tif, 02_120136E-1214-1%AR188-4-700.tif, 02_120136F-1206-1%AQ231-1-700.tif Chip Pfizer_120136 To do mouse serum binding: 1) 1:100 diluted mouse serum in 0.5 mL binding buffer, 4 ºC, 16 hours. 2) Wash with 1 mL of 1×PBS, pH7.0. 3) 10 ng/mL anti-mouse IgG Cy5 conjugate in 0.5 mL PBS buffer, 4 ºC, 2 hours. 4) Wash with 1 mL of 1×PBS, pH7.0. 5) Image scanned at 635 nm and PMT 700. A=1%Normal B=1%AR172-1 C=1%AR172-3 D=1%AR188-2 E=1%AR188-4 F=1%AQ231-1

4. 01_120138-1212-block-700.tif12/12/2012 by AH Chip Pfizer_120138 To block chip surface: 1) Block with SuperBlock, 0.05% Tween-20, pH 7.0, 4 ºC overnight. 2) Wash with 1 mL of 1×PBS, pH7.0. 3) Image scanned at 635 nm and PMT 700. A B C D E F

5. 02_120138A-1221-1%AR154-3-700.tif, 02_120138B-1222-1%AR154-6-700.tif, 02_120138C-1221-1%AR146-1-700.tif 02_120138D-1221-1%AR150-1-700.tif, 02_120138E-1221-1%AR147-11-700.tif, 02_120138F-1222-1%AR151-5-700.tif Chip Pfizer_120138 To do mouse serum binding: 1) 1:100 diluted mouse serum in 0.5 mL binding buffer, 4 ºC, 16 hours. 2) Wash with 1 mL of 1×PBS, pH7.0. 3) 10 ng/mL anti-mouse IgG Cy5 conjugate in 0.5 mL PBS buffer, 4 ºC, 2 hours. 4) Wash with 1 mL of 1×PBS, pH7.0. 5) Image scanned at 635 nm and PMT 700. A=1%AR154-3 B=1%AR154-6 C=1%AR146-1 D=1%AR150-1 E=1%AR147-11 F=1%AR151-5