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Human MANF(Mesencephalic Astrocyte Derived Neurotrophic Factor) ELISA Kit
ELK3084
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48 T $320.00
96 T $458.00
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Overview

Product name: Human MANF(Mesencephalic Astrocyte Derived Neurotrophic Factor) ELISA Kit
Reactivity: Human
Alternative Names: ARMET; ARP; Arginine Rich Mutated In Early Stage Tumors; Arginine-rich protein
Assay Type: Sandwich
Sensitivity: 6.1 pg/mL
Standard: 1000 pg/mL
Detection Range: 15.63-1000 pg/mL
Sample Type: serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Assay Length: 3.5h
Research Area: Tumor immunity;Neuro science;
Uniprot ID: P55145
Test principle: 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 Human MANF. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human 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 Human 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 Human MANF in the samples is then determined by comparing the OD of the samples to the standard curve.

Standard curve

Concentration (pg/mL) OD Corrected OD
1000.00 1.951 1.856
500.00 1.577 1.482
250.00 1.194 1.099
125.00 0.827 0.732
62.50 0.614 0.519
31.25 0.305 0.210
15.63 0.119 0.024
0.00 0.095 0.000

Precision

Intra-assay Precision (Precision within an assay)：CV%<8%

Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.

Inter-assay Precision (Precision between assays)：CV%<10%

Three samples of known concentration were tested in forty separate assays to assess inter-assay precision.

Recovery

Matrices listed below were spiked with certain level of recombinant MANF and the recovery rates were calculated by comparing the measured value to the expected amount of MANF in samples.
Matrix Recovery range Average
serum(n=5) 83-98% 90%
EDTA plasma(n=5) 81-95% 88%
Heparin plasma(n=5) 88-102% 95%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of MANF and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Matrix 1:2 1:4 1:8 1:16
serum(n=5) 95-102% 93-101% 97-105% 95-103%
EDTA plasma(n=5) 98-105% 86-92% 87-93% 95-103%
Heparin plasma(n=5) 96-101% 85-92% 79-96% 89-101%