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Rat GRP78(Glucose Regulated Protein 78) ELISA Kit
ELK6834
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48 T $320.00
96 T $458.00
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OverviewPublications

Product name: Rat GRP78(Glucose Regulated Protein 78) ELISA Kit
Reactivity: Rat
Alternative Names: MIF2; BIP; GRP78; Immunoglobulin heavy chain-binding protein; Glucose Regulated Protein 78; Binding Immunoglobulin Protein; Endoplasmic reticulum lumenal Ca binding grp78; Heat Shock 70kDa Protein 5; HSPA5
Assay Type: Sandwich
Sensitivity: 0.31 ng/mL
Standard: 50 ng/mL
Detection Range: 0.79-50 ng/mL
Sample Type: Tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Assay Length: 3.5h
Research Area: Signal transduction;Tumor immunity;Infection immunity;
Uniprot ID: P06761
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 Rat GRP78. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Rat GRP78. 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 Rat GRP78, 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 Rat GRP78 in the samples is then determined by comparing the OD of the samples to the standard curve.

Standard curve

Concentration (ng/mL) OD Corrected OD
50.00 2.132 2.044
25.00 1.636 1.548
12.50 1.243 1.155
6.25 0.863 0.775
3.13 0.496 0.408
1.57 0.335 0.247
0.79 0.168 0.080
0.00 0.088 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 GRP78 and the recovery rates were calculated by comparing the measured value to the expected amount of GRP78 in samples.
Matrix Recovery range Average
serum(n=5) 80-95% 87%
EDTA plasma(n=5) 78-96% 87%
Heparin plasma(n=5) 78-92% 85%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of GRP78 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) 97-106% 86-92% 91-103% 89-97%
EDTA plasma(n=5) 85-97% 90-99% 82-101% 93-105%
Heparin plasma(n=5) 82-95% 88-102% 84-97% 89-107%