TNFSF11 Chemi-Luminescent ELISA Kit (Rat)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Receptor Activator Of Nuclear Factor Kappa B Ligand (RANkL) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Receptor Activator Of Nuclear Factor Kappa B Ligand (RANkL) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

ODF; OPGL; osteoclast differentiation factor; osteoprotegerin ligand; RANKL; receptor activator of nuclear factor kappa-B ligand; TNF-related activation-induced cytokine; TRANCE; tumor necrosis factor (ligand) superfamily, member 11; tumor necrosis factor ligand superfamily member 11; tumor necrosis factor superfamily member 11.

Cytokine that binds to TNFRSF11B/OPG and to TNFRSF11A/RANK. Osteoclast differentiation and activation factor. Augments the ability of dendritic cells to stimulate naive T-cell proliferation. May be an important regulator of interactions between T-cells and dendritic cells and may play a role in the regulation of the T-cell-dependent immune response. May also play an important role in enhanced bone-resorption in humoral hypercalcemia of malignancy. Induces osteoclastogenesis by activating multiple signaling pathways in osteoclast precursor cells, chief among which is induction of long lasting oscillations in the intracellular concentration of Ca (2+) resulting in the activation of NFATC1, which translocates to the nucleus and induces osteoclast-specific gene transcription to allow differentiation of osteoclasts. During osteoclast differentiation, in a TMEM64 and ATP2A2-dependent manner induces activation of CREB1 and mitochondrial ROS generation necessary for proper osteoclast generation.

The microplate provided in this kit has been pre-coated with an antibody specific to Receptor Activator Of Nuclear Factor Kappa B Ligand (RANkL) . Standards or samples are then added to the appropriate microplate wells with a biotin-conjugated antibody specific to Receptor Activator Of Nuclear Factor Kappa B Ligand (RANkL) . 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 Receptor Activator Of Nuclear Factor Kappa B Ligand (RANkL) 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) 82-90 86 EDTA Plasma (n=5) 78-101 83 Heparin Plasma (n=5) 94-102 97 Matrix Recovery range (%) Average (%) Matrix Recovery range (%) Average (%) Serum (n=5) 82-90 86 Serum (n=5) 82-90 86 EDTA Plasma (n=5) 78-101 83 EDTA Plasma (n=5) 78-101 83 Heparin Plasma (n=5) 94-102 97 Heparin Plasma (n=5) 94-102 97

Component Amount Tnfsf11 Microplate 96 Wells (12 x 8 Well strips) Tnfsf11 Lyophilized Standard 2 100X Tnfsf11 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 Tnfsf11 Microplate 96 Wells (12 x 8 Well strips) Tnfsf11 Microplate 96 Wells (12 x 8 Well strips) Tnfsf11 Lyophilized Standard 2 Tnfsf11 Lyophilized Standard 2 100X Tnfsf11 Biotinylated Detector Anitbody 1 x 120 uL 100X Tnfsf11 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