Human CCL21/6Ckine ELISA Kit EZ-SetTM (DIY Antibody Pairs)

Chemokine (C-C motif) ligand 21 (CCL21) is a small cytokine belonging to the CC chemokine family. This chemokine is also known as 6Ckine (because it has six conserved cysteine residues instead of the four cysteines typical to chemokines), exodus-2, and secondary lymphoid-tissue chemokine (SLC) . By somatic cell hybrid and radiation hybrid analyses, mapped the SCYA21 gene to 9p13. Chemokines are a family of proteins that leukocyte migration and activation to inflammatory stimuli. CXC chemokine ligand 13 (CXCL13), CC chemokine ligand 21 (CCL21), and CCL19 are constitutively expressed in secondary lymphoid organs, where they control the placement of lymphocytes and dendritic cells. It was demonstrate that the local expression of homeostatic chemokines in nonlymphoid organs, such as the lung, plays an important role in protective immune responses.

C-C motif chemokine 21; 6Ckine; Beta-chemokine exodus-2; Secondary lymphoid-tissue chemokine; SLC; Small-inducible cytokine A21; CCL21; SCYA21; UNQ784/PRO1600

Highly expressed in high endothelial venules of lymph nodes, spleen and appendix. Intermediate levels found in small intestine, thyroid gland and trachea. Low level expression in thymus, bone marrow, liver, and pancreas. Also found in tonsil, fetal heart and fetal spleen.

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 100 709 1352 103 706 1475 Standard deviation 5.6 28.36 82.47 6.9 31.77 92.92 CV (%) 5.6% 4% 6.1% 6.7% 4.5% 6.3% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 100 709 1352 103 706 1475 Standard deviation 5.6 28.36 82.47 6.9 31.77 92.92 CV (%) 5.6% 4% 6.1% 6.7% 4.5% 6.3% 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) 100 709 1352 103 706 1475 Mean (pg/ml) 100 709 1352 103 706 1475 Standard deviation 5.6 28.36 82.47 6.9 31.77 92.92 Standard deviation 5.6 28.36 82.47 6.9 31.77 92.92 CV (%) 5.6% 4% 6.1% 6.7% 4.5% 6.3% CV (%) 5.6% 4% 6.1% 6.7% 4.5% 6.3%

Inhibits hemopoiesis and stimulates chemotaxis. Chemotactic in vitro for thymocytes and activated T-cells, but not for B-cells, macrophages, or neutrophils. Shows preferential activity towards naive T-cells. May play a role in mediating homing of lymphocytes to secondary lymphoid organs. Binds to atypical chemokine receptor ACKR4 and mediates the recruitment of beta-arrestin (ARRB1/2) to ACKR4.

Catalog number Description Quantity Storage of opened/reconstituted material EZ0555-CA Rabbit anti- human CCL21 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. EZ0555-DA Biotinylated goat anti- human CCL21 polyclonal antibody (Detection Antibody) 500 μl, 10 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0555-ST Lyophilized recombinant human CCL21 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 EZ0555-CA Rabbit anti- human CCL21 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. EZ0555-DA Biotinylated goat anti- human CCL21 polyclonal antibody (Detection Antibody) 500 μl, 10 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0555-ST Lyophilized recombinant human CCL21 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 EZ0555-CA Rabbit anti- human CCL21 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. EZ0555-CA Rabbit anti- human CCL21 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. EZ0555-DA Biotinylated goat anti- human CCL21 polyclonal antibody (Detection Antibody) 500 μl, 10 μg/mL (recommended dilution 1:100) EZ0555-DA Biotinylated goat anti- human CCL21 polyclonal antibody (Detection Antibody) 500 μl, 10 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) EK0555-ST Lyophilized recombinant human CCL21 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. EK0555-ST Lyophilized recombinant human CCL21 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 62.5 125 250 500 1000 2000 4000 O.D. 0.031 0.057 0.102 0.203 0.408 0.504 1.178 2.318 Concentration (pg/ml) 0 62.5 125 250 500 1000 2000 4000 O.D. 0.031 0.057 0.102 0.203 0.408 0.504 1.178 2.318 Concentration (pg/ml) 0 62.5 125 250 500 1000 2000 4000 Concentration (pg/ml) 0 62.5 125 250 500 1000 2000 4000 O.D. 0.031 0.057 0.102 0.203 0.408 0.504 1.178 2.318 O.D. 0.031 0.057 0.102 0.203 0.408 0.504 1.178 2.318

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)