Rat MIP-1Alpha/CCL3 ELISA Kit EZ-SetTM (DIY Antibody Pairs)

Macrophage inflammatory protein-1 alpha (MIP-1 alpha), also called CCL3, LD78. The cDNA for MIP-1 alpha predicts a mature peptide of 69 amino acids with a molecular mass of 7,889 daltons. LD78 is a member of a newly identified superfamily of small inducible proteins involved in inflammatory responses, wound healing and tumorigenesis. MIP-1 alpha is a chemokine that has pro-inflammatory and stem cell inhibitory activities in vitro. It constitutes an important second signal for mast cell degranulation in the conjunctiva in vivo and consequently for acute-phase disease.

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 53 177 293 65 192 317 Standard deviation 2.76 11.86 13.2 4.1 14.6 18.4 CV (%) 5.2% 6.7% 4.5% 6.3% 7.6% 5.8% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 53 177 293 65 192 317 Standard deviation 2.76 11.86 13.2 4.1 14.6 18.4 CV (%) 5.2% 6.7% 4.5% 6.3% 7.6% 5.8% 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) 53 177 293 65 192 317 Mean (pg/ml) 53 177 293 65 192 317 Standard deviation 2.76 11.86 13.2 4.1 14.6 18.4 Standard deviation 2.76 11.86 13.2 4.1 14.6 18.4 CV (%) 5.2% 6.7% 4.5% 6.3% 7.6% 5.8% CV (%) 5.2% 6.7% 4.5% 6.3% 7.6% 5.8%

Monokine with inflammatory and chemokinetic properties. Has chemotactic activity for monocytes, neutrophils, eosinophils, basophils, and lymphocytes. Required for lung TNF-alpha production, neutrophil recruitment and subsequent lung injury and may function as an autocrine mediator for the macrophage production of TNF-alpha which in turn up-regulates vascular adhesion molecules required for neutrophil influx. This protein binds heparin.

Catalog number Description Quantity Storage of opened/reconstituted material EZ1219-CA Rabbit anti- rat Ccl3 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. EZ1219-DA Biotinylated goat anti- rat Ccl3 polyclonal antibody (Detection Antibody) 500 μl, 3.3 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK1219-ST Lyophilized recombinant rat Ccl3 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 EZ1219-CA Rabbit anti- rat Ccl3 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. EZ1219-DA Biotinylated goat anti- rat Ccl3 polyclonal antibody (Detection Antibody) 500 μl, 3.3 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK1219-ST Lyophilized recombinant rat Ccl3 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 EZ1219-CA Rabbit anti- rat Ccl3 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. EZ1219-CA Rabbit anti- rat Ccl3 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. EZ1219-DA Biotinylated goat anti- rat Ccl3 polyclonal antibody (Detection Antibody) 500 μl, 3.3 μg/mL (recommended dilution 1:100) EZ1219-DA Biotinylated goat anti- rat Ccl3 polyclonal antibody (Detection Antibody) 500 μl, 3.3 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) EK1219-ST Lyophilized recombinant rat Ccl3 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. EK1219-ST Lyophilized recombinant rat Ccl3 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 7.8 15.6 31.3 62.5 125 250 500 O.D. 0.137 0.218 0.283 0.422 0.691 1.101 1.816 2.423 Concentration (pg/ml) 0 7.8 15.6 31.3 62.5 125 250 500 O.D. 0.137 0.218 0.283 0.422 0.691 1.101 1.816 2.423 Concentration (pg/ml) 0 7.8 15.6 31.3 62.5 125 250 500 Concentration (pg/ml) 0 7.8 15.6 31.3 62.5 125 250 500 O.D. 0.137 0.218 0.283 0.422 0.691 1.101 1.816 2.423 O.D. 0.137 0.218 0.283 0.422 0.691 1.101 1.816 2.423

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)