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Human HMGB-1(High mobility group protein B1)ELISA Kit (HUES02568)

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SKU:
HUES02568
$613

Description

Human HMGB-1(High mobility group protein B1)ELISA Kit

This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human HMGB-1 . Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human HMGB-1 and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human HMGB-1, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human HMGB-1. The concentration of Human HMGB-1 in samples can be calculated by comparing the OD of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityHuman
Detection MethodColormetric
Detection Range31.25-2000 pg/mL
Sensitivity18.75 pg/mL
Sample Volume Required Per Well100uL
Sample TypeSerum, plasma and other biological fluids

Specificity

This kit recognizes Human HMGB-1 in samples. No significant cross-reactivity or interference between Human HMGB-1 and analogues was observed.

Typical Data

As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

Concentration
(pg/mL)
O.D Average Corrected
20002.443
2.447
2.4452.374
10001.718
1.764
1.7411.67
5000.988
0.984
0.9860.915
2500.466
0.498
0.4820.411
1250.265
0.247
0.2560.185
62.50.187
0.161
0.1740.103
31.250.118
0.13
0.1240.053
00.069
0.073
0.071--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human HMGB-1 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human HMGB-1 were tested on 3 different plates, 20 replicates in each plate.

Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)99.32202.75693.26108.96208.03746.59
Standard deviation5.848.6235.565.869.7626.06
C V (%)5.884.255.135.384.693.49

Recovery

The recovery of Human HMGB-1 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type Range (%) Average Recovery (%)
Serum (n=5)94-108100
EDTA plasma (n=5)94-107100
Cell culture media (n=5)93-10599

Linearity

Samples were spiked with high concentrations of Human HMGB-1 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
1:2Range (%)98-11192-10585-100
Average (%)1059892
1:4Range (%)90-10384-10088-102
Average (%)989194
1:8Range (%)87-9983-9385-96
Average (%)948890
1:16Range (%)88-10385-9686-99
Average (%)949091

Kit Components & Storage

An unopened kit can be stored at 4'C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

Item Specifications Storage
Micro ELISA Plate(Dismountable)8 wells ×12 strips-20'C, 6 months
Reference Standard2 vials
Concentrated Biotinylated Detection Ab (100×)1 vial, 120 uL
Concentrated HRP Conjugate (100×)1 vial, 120 uL-20'C(shading light), 6 months
Reference Standard & Sample Diluent1 vial, 20 mL4'C, 6 months
Biotinylated Detection Ab Diluent1 vial, 14 mL
HRP Conjugate Diluent1 vial, 14 mL
Concentrated Wash Buffer (25×)1 vial, 30 mL
Substrate Reagent1 vial, 10 mL4'C(shading light)
Stop Solution1 vial, 10 mL4'C
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 copy
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Additional Information

Reactivity:
Human
ELISA Type:
Sandwich
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