Products

Rat HMGB-1(High Mobility Group Protein B1) ELISA Kit
ELK10272
Size： Price：
48 T $320.00
96 T $458.00
Manual MSDS Send email to us

Overview

Product name: Rat HMGB-1(High Mobility Group Protein B1) ELISA Kit
Reactivity: Rat
Alternative Names: HMGB1; HMG3; SBP1; Sulfoglucuronyl Carbohydrate Binding Protein; Amphoterin; High Mobility Group Box 1 Protein; HMG1; High Mobility Group Protein 1
Assay Type: Sandwich
Sensitivity: 0.067 ng/mL
Standard: 10 ng/mL
Detection Range: 0.16-10 ng/mL
Sample Type: serum, plasma, tissue homogenates and other biological fluids
Assay Length: 3.5h
Research Area: Infection immunity;
Uniprot ID: P63159
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 HMGB-1. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Rat HMGB-1. 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 HMGB-1, 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 HMGB-1 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
10.00 1.978 1.892
5.00 1.576 1.490
2.50 1.118 1.032
1.25 0.857 0.771
0.63 0.496 0.410
0.32 0.372 0.286
0.16 0.187 0.101
0.00 0.086 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 HMGB-1 and the recovery rates were calculated by comparing the measured value to the expected amount of HMGB-1 in samples.
Matrix Recovery range Average
serum(n=5) 96-107% 101%
EDTA plasma(n=5) 90-105% 97%
Heparin plasma(n=5) 85-97% 91%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of HMGB-1 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) 92-102% 81-93% 86-98% 91-103%
EDTA plasma(n=5) 85-97% 87-96% 85-96% 85-95%
Heparin plasma(n=5) 86-95% 93-103% 87-98% 85-96%