Products

Human Anti-GAD65(Anti-Glutamate Decarboxylase 65 Autoantibodies) ELISA Kit
ELK0506
Size： Price：
48 T $320.00
96 T $458.00
Download ① Send email to us

Overview

Product name: Human Anti-GAD65(Anti-Glutamate Decarboxylase 65 Autoantibodies) ELISA Kit
Reactivity: Human
Alternative Names: anti-GAD65; anti-Pancreatic Islets And Brain,65kDa; anti-65 kDa glutamic acid decarboxylase; anti-Glutamate decarboxylase 65 kDa isoform
Assay Type: Sandwich
Sensitivity: 1.31 IU/mL
Standard: 250 IU/mL
Detection Range: 3.91-250 IU/mL
Sample Type: tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Assay Length: 3.5h
Research Area: Enzyme & Kinase;Endocrinology;Neuro science;Autoimmunity;
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 Anti-GAD65. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human Anti-GAD65. 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 Anti-GAD65, 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 Anti-GAD65 in the samples is then determined by comparing the OD of the samples to the standard curve.

Standard curve

Concentration (IU/mL) OD Corrected OD
250.00 2.096 2.014
125.00 1.613 1.531
62.50 1.174 1.092
31.25 0.782 0.700
15.63 0.574 0.492
7.82 0.371 0.289
3.91 0.249 0.167
0.00 0.082 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 Anti-GAD65 and the recovery rates were calculated by comparing the measured value to the expected amount of Anti-GAD65 in samples.
Matrix Recovery range Average
serum(n=5) 90-105% 97%
EDTA plasma(n=5) 88-102% 95%
Heparin plasma(n=5) 82-97% 89%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Anti-GAD65 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) 78-92% 83-96% 95-107% 87-102%
EDTA plasma(n=5) 95-104% 93-102% 79-94% 87-98%
Heparin plasma(n=5) 89-100% 79-91% 89-103% 82-95%