Human VCAM-1 ELISA Kit EZ-SetTM (DIY Antibody Pairs)

Vascular cell adhesion molecule 1 (VCAM-1) is a cell surface glycoprotein adhesive for certain blood leukocytes and tumor cells, which is expressed by activated endothelium in a variety of pathologic conditions including atherosclerosis. Increased expression of VCAM1 is associated with a variety of chronic inflammatory conditions, making its expression and function a target for therapeutic intervention. Integrin alpha4beta1 (VLA-4) and VCAM-1 facilitate a critical cell-cell adhesion event required for survival of endothelial and mural cells during vascularization. The VCAM1 gene is assigned to the 1p31-32 region of chromosome 1 based on the analysis of human-mouse hybrid cell lines and in situ hybridization. The standard product used in this kit is recombinant human VCAM-1, consisting of Phe25-Glu698 amino acid sequence of a single chain with the molecular mass of 74KDa. As a result of glycosylation, the molecular mass of 90-95KDa is revealed by SDS-PAGE.

Atherosclerosis, Cancer, Cardiovascular, Cell Adhesion, Cell Adhesion Molecules, Cytoskeleton/ECM, Invasion/Microenvironment, Mesenchymal Stem Cells, Neural Signal Transduction, Neurology Process, Neuroscience, Signal Transduction, Stem Cells, Surface Molecules, Vascular Inflammation

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 560 3210 6560 450 2860 5890 Standard deviation 24.08 160.5 229.6 26.1 185.9 282.72 CV (%) 4.3% 5% 3.5% 5.8% 6.5% 4.8% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 560 3210 6560 450 2860 5890 Standard deviation 24.08 160.5 229.6 26.1 185.9 282.72 CV (%) 4.3% 5% 3.5% 5.8% 6.5% 4.8% 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) 560 3210 6560 450 2860 5890 Mean (pg/ml) 560 3210 6560 450 2860 5890 Standard deviation 24.08 160.5 229.6 26.1 185.9 282.72 Standard deviation 24.08 160.5 229.6 26.1 185.9 282.72 CV (%) 4.3% 5% 3.5% 5.8% 6.5% 4.8% CV (%) 4.3% 5% 3.5% 5.8% 6.5% 4.8%

Important in cell-cell recognition. Appears to function in leukocyte-endothelial cell adhesion. Interacts with integrin alpha-4/beta-1 (ITGA4/ITGB1) on leukocytes, and mediates both adhesion and signal transduction. The VCAM1/ITGA4/ITGB1 interaction may play a pathophysiologic role both in immune responses and in leukocyte emigration to sites of inflammation.

Catalog number Description Quantity Storage of opened/reconstituted material EZ0537-CA Rabbit anti- human VCAM1 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0537-DA Biotinylated goat anti- human VCAM1 polyclonal antibody (Detection Antibody) 500 μl, 14 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0537-ST Lyophilized recombinant human VCAM1 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material EZ0537-CA Rabbit anti- human VCAM1 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0537-DA Biotinylated goat anti- human VCAM1 polyclonal antibody (Detection Antibody) 500 μl, 14 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0537-ST Lyophilized recombinant human VCAM1 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material Catalog number Description Quantity Storage of opened/reconstituted material EZ0537-CA Rabbit anti- human VCAM1 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0537-CA Rabbit anti- human VCAM1 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0537-DA Biotinylated goat anti- human VCAM1 polyclonal antibody (Detection Antibody) 500 μl, 14 μg/mL (recommended dilution 1:100) EZ0537-DA Biotinylated goat anti- human VCAM1 polyclonal antibody (Detection Antibody) 500 μl, 14 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0537-ST Lyophilized recombinant human VCAM1 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. EK0537-ST Lyophilized recombinant human VCAM1 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit.

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.033 0.157 0.245 0.475 0.844 1.293 1.735 2.365 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.033 0.157 0.245 0.475 0.844 1.293 1.735 2.365 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.033 0.157 0.245 0.475 0.844 1.293 1.735 2.365 O.D. 0.033 0.157 0.245 0.475 0.844 1.293 1.735 2.365

1. Microplate reader in standard size. 2. Automated plate washer. 3. Incubator. 4. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. 5. Clean tubes and Eppendorf tubes. 6. 96 well microplate (Cat# AR1100) 7. Plate Sealers. 8. Capture Antibody Diluent: PBS. 9. Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 μm filtered. 10. Substrate Solution: Tetramethylbenzidine (Cat# AR1104) 11. Stop Solution: 2 N H2SO4 (Cat# AR1105) 12. Wash Buffer (PBS and PBS-T) . Recipes: PBS: 8g NaCl, 0.2g KCl, 1.15g Na2HPO4, 0.2g KH2PO4, adjust the total volume to 1 L with distilled water, pH 7.2-7.4, 0.2 μm filtered. PBS-T: 0.1% Tween® 20 in PBS, pH 7.2-7.4. *Item 6 – 12 are included in the EZ Set Accessory Kit (EZA001)