Mouse PECAM-1/CD31/PECAM1 PicoKine® Quick ELISA Kit

PECAM-1 is an important target GP in DITP.1 Platelet/endothelial cell adhesion molecule-1 (PECAM-1) is a 130-kD member of the Ig gene superfamily that is expressed on the surface of circulating platelets, monocytes, neutrophils, and selective T-cell subsets.2 Platelet endothelial cell adhesion molecule-1 (PECAM-1/CD31) is a member of the immunoglobulin (Ig) superfamily that has distinctive features of an immunoreceptor based upon its genomic structure and the presence of intrinsic immunoreceptor tyrosine inhibitory motifs (ITIMs) in its ligand binding polypeptide.3 PECAM-1 has recently been shown to contain functional immunoreceptor tyrosine-based inhibitory motifs (ITIMs) within its cytoplasmic domain, and co-ligation of PECAM-1 with the T-cell antigen receptor (TCR) results in tyrosine phosphorylation of PECAM-1, recruitment of Src homology 2 domain-containing protein tyrosine phosphatase-2 (SHP-2), and attenuation of TCR-mediated cellular signaling.4 The standard used in this kit is the product of gene recombinant expression (extracellular part), consisting of 563 amino acids (E28-K590) with the molecular weight of 65.5KDa. After glycosylating, its molecular weight changes to 91-100KDa.

Reproducibility To assay reproducibility, three samples with differing target protein concentrations were assayed using four different lots. Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 230 218 233 197 217 12.59 5.8% Sample 2 1352 1365 1136 1340 1298 66.21 5.1% Sample 3 5614 5784 5696 6014 5777 202.2 3.5% Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 230 218 233 197 217 12.59 5.8% Sample 1 230 218 233 197 217 12.59 5.8% Sample 2 1352 1365 1136 1340 1298 66.21 5.1% Sample 2 1352 1365 1136 1340 1298 66.21 5.1% Sample 3 5614 5784 5696 6014 5777 202.2 3.5% Sample 3 5614 5784 5696 6014 5777 202.2 3.5%

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 230 1352 5614 236 1369 5244 Standard deviation 17.94 73.01 387.37 18.41 97.2 361.84 CV (%) 7.8% 5.4% 6.9% 7.8% 7.1% 6.9% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 230 1352 5614 236 1369 5244 Standard deviation 17.94 73.01 387.37 18.41 97.2 361.84 CV (%) 7.8% 5.4% 6.9% 7.8% 7.1% 6.9% Intra-Assay Precision Inter-Assay Precision Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 n 16 16 16 24 24 24 Mean (pg/ml) 230 1352 5614 236 1369 5244 Mean (pg/ml) 230 1352 5614 236 1369 5244 Standard deviation 17.94 73.01 387.37 18.41 97.2 361.84 Standard deviation 17.94 73.01 387.37 18.41 97.2 361.84 CV (%) 7.8% 5.4% 6.9% 7.8% 7.1% 6.9% CV (%) 7.8% 5.4% 6.9% 7.8% 7.1% 6.9%

Description|Quantity|Volume Anti-tag Pre-coated 96-well Strip Microplate|1|12 strips of 8 wells Standard||2 Antibody A|1|3ml Antibody B|1|3ml Sample Diluent|1|15ml TBS-T Wash Buffer (25x) |1|12ml Color Developing Reagent (TMB) |1|10ml Stop Solution|1|10ml Adhesive Plate Sealers|2|Piece

Concentration (pg/ml) 0 156 312 625 1,250 2,500 5,000 10,000 O.D. 0.043 0.136 0.192 0.312 0.545 0.908 1.441 2.033 Concentration (pg/ml) 0 156 312 625 1,250 2,500 5,000 10,000 O.D. 0.043 0.136 0.192 0.312 0.545 0.908 1.441 2.033 Concentration (pg/ml) 0 156 312 625 1,250 2,500 5,000 10,000 Concentration (pg/ml) 0 156 312 625 1,250 2,500 5,000 10,000 O.D. 0.043 0.136 0.192 0.312 0.545 0.908 1.441 2.033 O.D. 0.043 0.136 0.192 0.312 0.545 0.908 1.441 2.033

Microplate Reader capable of reading absorbance at 450nm. Automated plate washer (optional) Pipettes and pipette tips capable of precisely dispensing 0.5 μl through 1 ml volumes of aqueous solutions. Multichannel pipettes are recommended for large amount of samples. Deionized or distilled water. 500ml graduated cylinders. Test tubes for dilution. Horizontal orbital microplate shaker capable of maintaining a speed of 500 rpm, amplitude 3mm.