Mouse CCL20/MIP-3 alpha ELISA Kit EZ-SetTM (DIY Antibody Pairs)

Macrophage Inflammatory Protein 3alpha (MIP3alpha), also called Chemokine, cc motif, ligand 20 (CCL20) . The MIP-3alpha/CCL20 gene was cloned and sequenced, revealing a four exon, three intron structure, and was localized by FISH analysis to 2q35-q36. MIP3alpha is predominantly expressed in lymph nodes, appendix, PBL, fetal liver, fetal lung and several cell lines. MIP3alpha/CCL20 and its receptor CCR6 are markedly up-regulated in psoriasis, and they may play a role in the recruitment of T cells to lesional psoriatic skin. And Alanine MIP-3alpha and Serine MIP-3alpha, the two forms of MIP3alpha, that differ by one amino acid at the predicted signal peptide cleavage site. Both of them were chemically synthesized and tested for biological activity. And both flu antigen plus IL-2-activated CD4 (+) and CD8 (+) T lymphoblasts and cord blood-derived dendritic cells responded to Ser and Ala MIP-3alpha.

C-C motif chemokine 20; Beta-chemokine exodus-1; CC chemokine LARC; CC chemokine ST38; Liver and activation-regulated chemokine; Macrophage inflammatory protein 3 alpha; MIP-3-alpha; Small-inducible cytokine A20; Ccl20; Larc; Scya20

Thymic medulla (at protein level) . Prominently expressed in the small intestine, colon and appendix. Also found in thymus, spleen, lymph node and lung. The long form might be dominant in intestinal, and the short form in lymphoid tissues. Expressed by IL17 producing helper T-cells (Th17) .

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 9 55 313 9 57 308 Standard deviation 0.62 3.02 21.28 0.78 3.7 21.56 CV (%) 6.9% 5.5% 6.8% 8.7% 6.5% 7% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 9 55 313 9 57 308 Standard deviation 0.62 3.02 21.28 0.78 3.7 21.56 CV (%) 6.9% 5.5% 6.8% 8.7% 6.5% 7% 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) 9 55 313 9 57 308 Mean (pg/ml) 9 55 313 9 57 308 Standard deviation 0.62 3.02 21.28 0.78 3.7 21.56 Standard deviation 0.62 3.02 21.28 0.78 3.7 21.56 CV (%) 6.9% 5.5% 6.8% 8.7% 6.5% 7% CV (%) 6.9% 5.5% 6.8% 8.7% 6.5% 7%

Acts as a ligand for C-C chemokine receptor CCR6. Signals through binding and activation of CCR6 and induces a strong chemotactic response and mobilization of intracellular calcium ions. The ligand-receptor pair CCL20-CCR6 is responsible for the chemotaxis of dendritic cells (DC), effector/memory T-cells and B-cells and plays an important role at skin and mucosal surfaces under homeostatic and inflammatory conditions, as well as in pathology, including cancer and autoimmune diseases. CCL20 acts as a chemotactic factor that attracts lymphocytes and, slightly, neutrophils, but not monocytes (By similarity) . Involved in the recruitment of both the proinflammatory IL17 producing helper T-cells (Th17) and the regulatory T-cells (Treg) to sites of inflammation. Required for optimal migration of thymic natural regulatory T cells (nTregs) and DN1 early thymocyte progenitor cells. Positively regulates sperm motility and chemotaxis via its binding to CCR6 which triggers Ca2+ mobilization in the sperm which is important for its motility. May be involved in formation and function of the mucosal lymphoid tissues by attracting lymphocytes and dendritic cells towards epithelial cells.

Catalog number Description Quantity Storage of opened/reconstituted material EZ0454-CA Rabbit anti- mouse Ccl20 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. EZ0454-DA Biotinylated goat anti- mouse Ccl20 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) EK0454-ST Lyophilized recombinant mouse Ccl20 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 EZ0454-CA Rabbit anti- mouse Ccl20 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. EZ0454-DA Biotinylated goat anti- mouse Ccl20 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) EK0454-ST Lyophilized recombinant mouse Ccl20 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 EZ0454-CA Rabbit anti- mouse Ccl20 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. EZ0454-CA Rabbit anti- mouse Ccl20 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. EZ0454-DA Biotinylated goat anti- mouse Ccl20 polyclonal antibody (Detection Antibody) 500 μl, 3.3 μg/mL (recommended dilution 1:100) EZ0454-DA Biotinylated goat anti- mouse Ccl20 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) EK0454-ST Lyophilized recombinant mouse Ccl20 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. EK0454-ST Lyophilized recombinant mouse Ccl20 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. 0.001 0.09 0.183 0.357 0.644 1.136 1.78 2.179 Concentration (pg/ml) 0 7.8 15.6 31.3 62.5 125 250 500 O.D. 0.001 0.09 0.183 0.357 0.644 1.136 1.78 2.179 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. 0.001 0.09 0.183 0.357 0.644 1.136 1.78 2.179 O.D. 0.001 0.09 0.183 0.357 0.644 1.136 1.78 2.179

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)