CCL20 Chemi-Luminescent ELISA Kit (Rat)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Macrophage Inflammatory Protein 3 Alpha (MIP3a) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Macrophage Inflammatory Protein 3 Alpha (MIP3a) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

Beta-chemokine exodus-1; CC chemokine LARC; CC chemokine ST38; C-C motif chemokine 20; chemokine (C-C motif) ligand 20; liver and activation-regulated chemokine; macrophage inflammatory protein 3 alpha; MIP-3-alpha; Scya20; small inducible cytokine subfamily A20; small-inducible cytokine A20; ST38.

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. 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.

The microplate provided in this kit has been pre-coated with an antibody specific to Macrophage Inflammatory Protein 3 Alpha (MIP3a) . Standards or samples are then added to the appropriate microplate wells with a biotin-conjugated antibody specific to Macrophage Inflammatory Protein 3 Alpha (MIP3a) . Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then the mixture of substrate A and B is added to generate glow light emission kinetics. Upon plate development, the intensity of the emitted light is proportional to the Macrophage Inflammatory Protein 3 Alpha (MIP3a) level in the sample or standard.

Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol

Matrix Recovery range (%) Average (%) Serum (n=5) 82-99 89 EDTA Plasma (n=5) 80-90 85 Heparin Plasma (n=5) 92-99 96 Matrix Recovery range (%) Average (%) Matrix Recovery range (%) Average (%) Serum (n=5) 82-99 89 Serum (n=5) 82-99 89 EDTA Plasma (n=5) 80-90 85 EDTA Plasma (n=5) 80-90 85 Heparin Plasma (n=5) 92-99 96 Heparin Plasma (n=5) 92-99 96

Component Amount Ccl20 Microplate 96 Wells (12 x 8 Well strips) Ccl20 Lyophilized Standard 2 100X Ccl20 Biotinylated Detector Anitbody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 100X Luminol Substrate 1 x 2 mL Substrate Diluent 1 x 20 mL Component Amount Component Amount Ccl20 Microplate 96 Wells (12 x 8 Well strips) Ccl20 Microplate 96 Wells (12 x 8 Well strips) Ccl20 Lyophilized Standard 2 Ccl20 Lyophilized Standard 2 100X Ccl20 Biotinylated Detector Anitbody 1 x 120 uL 100X Ccl20 Biotinylated Detector Anitbody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 30X Wash Buffer 1 x 20 mL 100X Luminol Substrate 1 x 2 mL 100X Luminol Substrate 1 x 2 mL Substrate Diluent 1 x 20 mL Substrate Diluent 1 x 20 mL