Universal ELISA Kits

LTB4(Leukotriene B4) ELISA Kit
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Product No

FY-EU1159

Sample Type

Serum, Plasma, Tissue homogenates, Cell culture supernates and Other biological fluids

Assay Time

1.5H

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Product Details
Product Name in English:
LTB4(Leukotriene B4) ELISA Kit
Species:
Universal
Alternative Name:
LT-B4
Assay Type:
Competitive
Standard:
20000 pg/mL
Sensitivity:
89.3 pg/mL
Detection Range:
312.5-20000 pg/mL
Assay Time:
1.5H
Sample Type:
Serum, Plasma, Tissue homogenates, Cell culture supernates and Other biological fluids
Research:
Metabolic pathway;Infection immunity;Endocrinology;Hormone metabolism;
Test Principle:
This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with LTB4 protein. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to LTB4. Next,Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added. The enzyme-substrate reaction is terminated by the addition of stop solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of LTB4 in the samples is then determined 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
20000 0.188 0.19 0.189
10000 0.324 0.326 0.325
5000 0.539 0.543 0.541
2500 0.873 0.881 0.877
1250 1.301 1.302 1.302
625 1.693 1.708 1.701
312.5 2.069 2.079 2.074
0 2.509 2.518 2.514
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) 89-101 95
EDTA plasma (n=8) 82-95 88
Cell culture media (n=8) 80-94 87
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 (%) 87-98 93-102 82-95
Average (%) 92 96 86
1:4 Range (%) 92-101 93-102 88-103
Average (%) 96 94 92
1:8 Range (%) 82-98 81-96 86-97
Average (%) 92 82 93
1:16 Range (%) 89-97 93-106 87-102
Average (%) 94 97 92
Assay Procedures
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