MANF ELISA Kit (Mouse)

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

3230402M22Rik; AA407711; AA408789; AA673178; arginine-rich protein; arginine-rich, mutated in early stage tumors; Arm; Armet; D17914; D18Mgi17; mesencephalic astrocyte-derived neurotrophic factor; Protein ARMET.

Selectively promotes the survival of dopaminergic neurons of the ventral mid-brain. Modulates GABAergic transmission to the dopaminergic neurons of the substantia nigra. Enhances spontaneous, as well as evoked, GABAergic inhibitory postsynaptic currents in dopaminergic neurons. Inhibits cell proliferation and endoplasmic reticulum (ER) stress-induced cell death. Retained in the ER/sarcoplasmic reticulum (SR) through association with the endoplasmic reticulum chaperone protein HSPA5 under normal conditions. Up-regulated and secreted by the ER/SR in response to ER stress and hypoxia. Following secretion by the ER/SR, directly binds to 3-O-sulfogalactosylceramide, a lipid sulfatide in the outer cell membrane of target cells. Sulfatide binding promotes its cellular uptake by endocytosis, and is required for its role in alleviating ER stress and cell toxicity under hypoxic and ER stress conditions.

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 Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF) . 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 Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF), 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 Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF) 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) 78-97 92 EDTA Plasma (n=5) 82-98 91 Heparin Plasma (n=5) 83-105 92 Matrix Recovery range (%) Average (%) Matrix Recovery range (%) Average (%) Serum (n=5) 78-97 92 Serum (n=5) 78-97 92 EDTA Plasma (n=5) 82-98 91 EDTA Plasma (n=5) 82-98 91 Heparin Plasma (n=5) 83-105 92 Heparin Plasma (n=5) 83-105 92

Component Amount Anti-MANF Microplate 96 Wells (12 x 8 Well strips) MANF Lyophilized Standard 2 100X Biotinylated MANF 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-MANF Microplate 96 Wells (12 x 8 Well strips) Anti-MANF Microplate 96 Wells (12 x 8 Well strips) MANF Lyophilized Standard 2 MANF Lyophilized Standard 2 100X Biotinylated MANF Detector Antibody 1 x 120 uL 100X Biotinylated MANF 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 Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF) ._x005F No significant cross-reactivity or interference between Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF) and analogues was observed.