EasyPro Series ELISA Kits
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Manual(EN) 
Product Details
Product Name in English:
EasyPro Human SEMA3A(Semaphorin 3A) ELISA Kit
Species:
Human
Alternative Name:
SEMA3A, COLL1, HH16, Hsema-I, Hsema-III, SEMA1, SEMAD, SEMAIII, SEMAL, SemD, coll-1, semaphorin 3A
Uniport ID:
Q14563
Assay Type:
Sandwich
Standard:
10ng/mL
Sensitivity:
0.10 ng/mL
Detection Range:
0.16-10ng/mL
Assay Time:
1.5H
Sample Type:
Serum, Plasma, Tissue homogenate and Other biological samples；Sample Volume=50uL
Research:
Neuroscience，Cancer，Immunology
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 SEMA3A. Samples (or Standards) and and Horseradish Peroxidase (HRP) linked antibody specific for SEMA3A are added to the micro ELISA plate wells. SEMA3A in samples (or standards) combines with the coated antibody and HRP linked detection antibody special to SEMA3A. 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 SEMA3A. You can calculate the concentration of SEMA3A 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 |
|---|---|---|---|---|
| 10 | 2.41 | 2.44 | 2.425 | 2.373 |
| 5 | 1.725 | 1.755 | 1.74 | 1.688 |
| 2.5 | 1.009 | 1.021 | 1.015 | 0.963 |
| 1.25 | 0.502 | 0.509 | 0.506 | 0.454 |
| 0.63 | 0.263 | 0.264 | 0.264 | 0.212 |
| 0.32 | 0.14 | 0.143 | 0.142 | 0.09 |
| 0.16 | 0.093 | 0.094 | 0.094 | 0.042 |
| 0 | 0.052 | 0.052 | 0.052 | 0 |
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 (%) | 86-98 | 85-92 | 80-102 |
| Average (%) | 94 | 90 | 92 | |
| 1:4 | Range (%) | 83-96 | 85-92 | 95-107 |
| Average (%) | 95 | 87 | 103 | |
| 1:8 | Range (%) | 95-104 | 87-101 | 92-103 |
| Average (%) | 96 | 95 | 97 | |
| 1:16 | Range (%) | 81-95 | 88-102 | 81-99 |
| Average (%) | 85 | 90 | 97 |
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) | 87-99 | 93 |
| EDTA plasma (n=8) | 80-95 | 87 |
| Cell culture media (n=8) | 90-107 | 98 |
Assay Procedures
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