GDF5 Chemi-Luminescent ELISA Kit (Human)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Growth Differentiation Factor 5 (GDF5) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Growth Differentiation Factor 5 (GDF5) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

BDA1C; BMP14; BMP-14; bone morphogenetic protein 14; Cartilage-derived morphogenetic protein 1; cartilage-derived morphogenetic protein-1; CDMP1; DUPANS; growth/differentiation factor 5; LAP4; LAP-4; lipopolysaccharide-associated protein 4; LPS-associated protein 4; OS5; radotermin; SYM1B; SYNS2.

Growth factor involved in bone and cartilage formation. During cartilage development regulates differentiation of chondrogenic tissue through two pathways. Firstly, positively regulates differentiation of chondrogenic tissue through its binding of high affinity with BMPR1B and of less affinity with BMPR1A, leading to induction of SMAD1-SMAD5-SMAD8 complex phosphorylation and then SMAD protein signaling transduction (PubMed:24098149, PubMed:21976273, PubMed:15530414, PubMed:25092592) . Secondly, negatively regulates chondrogenic differentiation through its interaction with NOG (PubMed:21976273) . Required to prevent excessive muscle loss upon denervation. This function requires SMAD4 and is mediated by phosphorylated SMAD1/5/8 (By similarity) . Binds bacterial lipopolysaccharide (LPS) and mediates LPS-induced inflammatory response, including TNF secretion by monocytes (PubMed:11276205) .

The microplate provided in this kit has been pre-coated with an antibody specific to Growth Differentiation Factor 5 (GDF5) . Standards or samples are then added to the appropriate microplate wells with a biotin-conjugated antibody specific to Growth Differentiation Factor 5 (GDF5) . 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 Growth Differentiation Factor 5 (GDF5) 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) 80-95 82 EDTA Plasma (n=5) 95-103 99 Heparin Plasma (n=5) 80-99 91 Matrix Recovery range (%) Average (%) Matrix Recovery range (%) Average (%) Serum (n=5) 80-95 82 Serum (n=5) 80-95 82 EDTA Plasma (n=5) 95-103 99 EDTA Plasma (n=5) 95-103 99 Heparin Plasma (n=5) 80-99 91 Heparin Plasma (n=5) 80-99 91

Component Amount GDF5 Microplate 96 Wells (12 x 8 Well strips) GDF5 Lyophilized Standard 2 100X GDF5 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 GDF5 Microplate 96 Wells (12 x 8 Well strips) GDF5 Microplate 96 Wells (12 x 8 Well strips) GDF5 Lyophilized Standard 2 GDF5 Lyophilized Standard 2 100X GDF5 Biotinylated Detector Anitbody 1 x 120 uL 100X GDF5 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