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Catalog Number:RE10051
Trp(Tryptophan) ELISA Kit
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96 T

48 T

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Product Details
Product Name Trp(Tryptophan) ELISA Kit Species Universal
Uniprot ID N/A Alternative Names Tryptophan,2-Amino-3-(1H-indol-3-yl)propanoic acid
Detection method Competitive Sensitivity 0.54 μg/mL
Standard 100μg/mL Detection Range 1.56-100μg/mL
Sample type Serum, Plasma, Tissue homogenate and Other biological samples;Sample Volume=50μL
Reaction time 1.5H Research Area N/A
Test principle This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with Trp. Standards or samples are added to the appropriate micro ELISA plate wells then with a biotin-conjugated antibody specific to Trp. 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 sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Trp 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.

(μg/mL) OD
100.00 0.274
50.00 0.374
25.00 0.568
12.50 0.829
6.25 1.124
3.13 1.519
1.57 1.833
0.00 2.497
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.

Intra-assay Precision Inter-assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean(μg/mL) 5.66 13.60 33.02 6.14 14.43 35.95
Standard deviation 0.28 0.68 1.65 0.31 0.72 1.80
C V (%) 3.14 5.42 4.20 3.24 5.66 4.50
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) 88-103 95
EDTA plasma (n=8) 80-95 86
Cell culture media (n=8) 92-106 99
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 (%) 85-102 98-106 89-97
Average (%) 90 105 93
1:4 Range (%) 83-96 98-106 85-94
Average (%) 86 101 87
1:8 Range (%) 87-98 86-99 87-96
Average (%) 95 87 94
1:16 Range (%) 82-95 80-95 92-101
Average (%) 90 80 97
Assay Procedures