EasyPro Series ELISA Kits

EasyPro Human HMGB-1(High Mobility Group Protein B1) ELISA Kit
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Product No

FY-EH3059S

Sample Type

Serum, Plasma, Tissue homogenate and Other biological samples;Sample Volume=50uL

Assay Time

1.5H

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Product Details
Product Name in English:
EasyPro Human HMGB-1(High Mobility Group Protein B1) ELISA Kit
Species:
Human
Alternative Name:
HMGB1, HMG1, HMG3, SBP-1, Amphoterin
Uniport ID:
P09429
Assay Type:
Sandwich
Standard:
2000pg/mL
Sensitivity:
18.75 pg/mL
Detection Range:
31.25-2000pg/mL
Assay Time:
1.5H
Sample Type:
Serum, Plasma, Tissue homogenate and Other biological samples;Sample Volume=50uL
Research:
Metabolic pathway;Developmental science;Nutrition metabolism;Bone metabolism;
Test Principle:
This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to HMGB-1. Samples (or Standards) and and Horseradish Peroxidase (HRP) linked antibody specific for HMGB-1 are added to the micro ELISA plate wells. HMGB-1 in samples (or standards) combines with the coated antibody and HRP linked detection antibody special to HMGB-1. Excess conjugate and unbound sample or standard are washed from the plate. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm. The OD value is proportional to the concentration of HMGB-1. You can calculate the concentration of HMGB-1 in the samples by comparing the OD of the samples to the standard curve.
Technical 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 OD1 OD2 Average OD Corrected
2000 2.326 2.327 2.327 2.284
1000 1.532 1.552 1.542 1.499
500 0.901 0.913 0.907 0.864
250 0.418 0.424 0.421 0.378
125 0.239 0.242 0.241 0.198
62.5 0.139 0.141 0.14 0.097
31.25 0.092 0.094 0.093 0.05
0 0.042 0.043 0.043 0
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level were tested on 3 different plates, 20 replicates in each plate, respectively.
Item Intra-assay Precision Intra-assay Precision
Sample 3 3
Replicate 9 18
CV(%) 5 8
Rate of recovery
The recovery of 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=8) 96-109 102
EDTA plasma (n=8) 81-95 88
Cell culture media (n=8) 85-97 91
Linear
Samples were spiked with high concentrations of target proteins 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)
1:2 Range (%) 86-97 95-102 91-102
Average (%) 95 101 97
1:4 Range (%) 83-96 95-102 89-99
Average (%) 84 97 92
1:8 Range (%) 91-103 98-106 88-104
Average (%) 103 101 102
1:16 Range (%) 96-105 87-98 84-98
Average (%) 100 90 98
Assay Procedures
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