LGALS9 ELISA Kit (Rat)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Galectin 9 (GAL9) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Galectin 9 (GAL9) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

36 kDa beta-galactoside-binding lectin; gal-9; galectin-9; Lectin, galactose binding, soluble 9 (Galectin-9) ; lectin, galactoside-binding, soluble, 9; UAT; urate transporter/channel.

Binds galactosides (By similarity) . Has high affinity for the Forssman pentasaccharide (By similarity) . Ligand for HAVCR2/TIM3 (By similarity) . Binding to HAVCR2 induces T-helper type 1 lymphocyte (Th1) death (By similarity) . Also stimulates bactericidal activity in infected macrophages by causing macrophage activation and IL1B secretion which restricts intracellular bacterial growth (By similarity) . Ligand for P4HB; the interaction retains P4HB at the cell surface of Th2 T helper cells, increasing disulfide reductase activity at the plasma membrane, altering the plasma membrane redox state and enhancing cell migration (By similarity) . Ligand for CD44; the interaction enhances binding of SMAD3 to the FOXP3 promoter, leading to up-regulation of FOXP3 expression and increased induced regulatory T (iTreg) cell stability and suppressive function (By similarity) . Promotes ability of mesenchymal stromal cells to suppress T-cell proliferation (By similarity) . Expands regulatory T-cells and induces cytotoxic T-cell apoptosis following virus infection (By similarity) . Activates ERK1/2 phosphorylation inducing cytokine (IL-6, IL-8, IL-12) and chemokine (CCL2) production in mast and dendritic cells (By similarity) . Inhibits degranulation and induces apoptosis of mast cells (By similarity) . Induces maturation and migration of dendritic cells (By similarity) . Inhibits natural killer (NK) cell function (By similarity) . Can transform NK cell phenotype from peripheral to decidual during pregnancy (By similarity) . Astrocyte derived galectin-9 enhances microglial TNF production (PubMed:25158758) . May play a role in thymocyte-epithelial interactions relevant to the biology of the thymus. May provide the molecular basis for urate flux across cell membranes, allowing urate that is formed during purine metabolism to efflux from cells and serving as an electrogenic transporter that plays an important role in renal and gastrointestinal urate excretion (PubMed:8995305) . Highly selective to the anion urate (PubMed:8995305) .

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Galectin 9 (GAL9) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Galectin 9 (GAL9) . Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Galectin 9 (GAL9), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm +/- 10nm. The concentration of Galectin 9 (GAL9) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

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) 97-105 101 EDTA Plasma (n=5) 97-105 102 Heparin Plasma (n=5) 82-105 90 Matrix Recovery range (%) Average (%) Matrix Recovery range (%) Average (%) Serum (n=5) 97-105 101 Serum (n=5) 97-105 101 EDTA Plasma (n=5) 97-105 102 EDTA Plasma (n=5) 97-105 102 Heparin Plasma (n=5) 82-105 90 Heparin Plasma (n=5) 82-105 90

Component Amount Anti-LGALS9 Microplate 96 Wells (12 x 8 Well strips) LGALS9 Lyophilized Standard 2 100X Biotinylated LGALS9 Detector Antibody 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 Stop Solution 1 x 6 mL TMB Substrate 1 x 9 mL Component Amount Component Amount Anti-LGALS9 Microplate 96 Wells (12 x 8 Well strips) Anti-LGALS9 Microplate 96 Wells (12 x 8 Well strips) LGALS9 Lyophilized Standard 2 LGALS9 Lyophilized Standard 2 100X Biotinylated LGALS9 Detector Antibody 1 x 120 uL 100X Biotinylated LGALS9 Detector Antibody 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 Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL

This assay has high sensitivity and excellent specificity for detection of Galectin 9 (GAL9) . No significant cross-reactivity or interference between Galectin 9 (GAL9) and analogues was observed.