PEDF ELISA Kit (Cattle)

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

PEDF; pigment epithelium-derived factor; serine (or cysteine) proteinase inhibitor, clade F (alpha-2 antiplasmin, pigment epithelium derived factor), member 1; serpin F1.

Neurotrophic protein; induces extensive neuronal differentiation in retinoblastoma cells. Potent inhibitor of angiogenesis. As it does not undergo the S (stressed) to R (relaxed) conformational transition characteristic of active serpins, it exhibits no serine protease inhibitory activity.

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 Pigment Epithelium Derived Factor (PEDF) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Pigment Epithelium Derived Factor (PEDF) . 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 Pigment Epithelium Derived Factor (PEDF), 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 Pigment Epithelium Derived Factor (PEDF) 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) 96-103 101 EDTA Plasma (n=5) 79-90 83 Heparin Plasma (n=5) 88-101 93 Matrix Recovery range (%) Average (%) Matrix Recovery range (%) Average (%) Serum (n=5) 96-103 101 Serum (n=5) 96-103 101 EDTA Plasma (n=5) 79-90 83 EDTA Plasma (n=5) 79-90 83 Heparin Plasma (n=5) 88-101 93 Heparin Plasma (n=5) 88-101 93

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