Rat CXCL4/PF4 ELISA Kit EZ-SetTM (DIY Antibody Pairs)

Platelet factor 4 (PF4) is a small cytokine belonging to the CXC chemokine family that is also known as chemokine (C-X-C motif) ligand 4 (CXCL4) . The PF4 gene was localized on 4q12-q13. Chemokines play fundamental roles in the development, homeostasis, and function of the immune system, and they have effects on cells of the central nervous system as well as on endothelial cells involved in angiogenesis or angiostasis. Platelet factor-4 is a 70-amino acid protein that is released from the alpha-granules of activated platelets and binds with high affinity to heparin. Its major physiologic role appears to be neutralization of heparin-like molecules on the endothelial surface of blood vessels, thereby inhibiting local antithrombin III activity and promoting coagulation. As a strong chemoattractant for neutrophils and fibroblasts, PF4 probably has a role in inflammation and wound repair.

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 201 989 3125 233 1040 3397 Standard deviation 8.84 42.52 153.13 14.21 62.4 200.42 CV (%) 4.4% 4.3% 4.9% 6.1% 6% 5.9% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 201 989 3125 233 1040 3397 Standard deviation 8.84 42.52 153.13 14.21 62.4 200.42 CV (%) 4.4% 4.3% 4.9% 6.1% 6% 5.9% 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) 201 989 3125 233 1040 3397 Mean (pg/ml) 201 989 3125 233 1040 3397 Standard deviation 8.84 42.52 153.13 14.21 62.4 200.42 Standard deviation 8.84 42.52 153.13 14.21 62.4 200.42 CV (%) 4.4% 4.3% 4.9% 6.1% 6% 5.9% CV (%) 4.4% 4.3% 4.9% 6.1% 6% 5.9%

Released during platelet aggregation. Neutralizes the anticoagulant effect of heparin because it binds more strongly to heparin than to the chondroitin-4-sulfate chains of the carrier molecule. Chemotactic for neutrophils and monocytes. Inhibits endothelial cell proliferation.

Catalog number Description Quantity Storage of opened/reconstituted material EZ1651-CA Rabbit anti- rat Pf4 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. EZ1651-DA Biotinylated goat anti- rat Pf4 polyclonal antibody (Detection Antibody) 500 μl, 8 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK1651-ST Lyophilized recombinant rat Pf4 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 EZ1651-CA Rabbit anti- rat Pf4 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. EZ1651-DA Biotinylated goat anti- rat Pf4 polyclonal antibody (Detection Antibody) 500 μl, 8 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK1651-ST Lyophilized recombinant rat Pf4 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 EZ1651-CA Rabbit anti- rat Pf4 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. EZ1651-CA Rabbit anti- rat Pf4 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. EZ1651-DA Biotinylated goat anti- rat Pf4 polyclonal antibody (Detection Antibody) 500 μl, 8 μg/mL (recommended dilution 1:100) EZ1651-DA Biotinylated goat anti- rat Pf4 polyclonal antibody (Detection Antibody) 500 μl, 8 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) EK1651-ST Lyophilized recombinant rat Pf4 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. EK1651-ST Lyophilized recombinant rat Pf4 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. 0.055 0.129 0.218 0.361 0.619 1.002 1.6 2.356 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.055 0.129 0.218 0.361 0.619 1.002 1.6 2.356 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.055 0.129 0.218 0.361 0.619 1.002 1.6 2.356 O.D. 0.055 0.129 0.218 0.361 0.619 1.002 1.6 2.356

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)