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Human HSP-70/HSPA9(HeatShock Protein 70) ELISA Kit
ELK4514
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48 T $320.00
96 T $458.00
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Overview

Product name: Human HSP-70/HSPA9(HeatShock Protein 70) ELISA Kit
Reactivity: Human
Alternative Names: DnaK; 70 Kilodalton Heat Shock Proteins;Heat shock 70 kDa protein 1; 2; heat shock 70kDa protein 1A; HSP70; HSP70 1; HSP70 1; HSP70 2; HSP70 1A; HSP70.1; HSP70.2; HSP70I; HSP72; HSPA1; HSPA1A; HSPA1B; HSP70; Heat Shock Protein 70
Assay Type: Sandwich
Sensitivity: 1.33 ng/mL
Standard: 200 ng/mL
Detection Range: 3.13-200 ng/mL
Sample Type: serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Assay Length: 3.5h
Research Area: Tumor immunity;Infection immunity;Developmental science;
Uniprot ID: P38646
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 HSP-70/HSPA9. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human HSP-70/HSPA9. 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 HSP-70/HSPA9, 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 HSP-70/HSPA9 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
200.00 1.984 1.919
100.00 1.647 1.582
50.00 1.139 1.074
25.00 0.728 0.663
12.50 0.517 0.452
6.25 0.323 0.258
3.13 0.238 0.173
0.00 0.065 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 HSP-70/HSPA9 and the recovery rates were calculated by comparing the measured value to the expected amount of HSP-70/HSPA9 in samples.
Matrix Recovery range Average
serum(n=5) 82-98% 90%
EDTA plasma(n=5) 82-94% 88%
Heparin plasma(n=5) 80-97% 88%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of HSP-70/HSPA9 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) 85-97% 87-96% 85-96% 85-95%
EDTA plasma(n=5) 86-95% 93-103% 87-98% 85-96%
Heparin plasma(n=5) 92-102% 81-93% 86-98% 91-103%