|Detection range||0.31-20 ng/mL|
|Sample type &Sample volume||serum, plasma and other biological fluids; 100μL|
|Specificity||This kit recognizes Human SEMA4D in samples. No significant cross-reactivity or interference between Human SEMA4D and analogues was observed.|
|Reproducibility||Both intra-CV and inter-CV are < 10%.|
|Application||This ELISA kit applies to the in vitro quantitative determination of Human SEMA4D concentrations in serum, plasma and other biological fluids.|
|Micro ELISA Plate(Dismountable)||96T: 8 wells ×12 strips
48T: 8 wells ×6 strips
|-20℃, 6 months|
|Reference Standard||96T: 2 vials
48T: 1 vial
|Concentrated Biotinylated Detection Ab (100×)||96T: 1 vial, 120 μL
48T: 1 vial, 60 μL
|Concentrated HRP Conjugate (100×)||96T: 1 vial, 120 μL
48T: 1 vial, 60 μL
|-20℃(Protect from light), 6 months|
|Reference Standard & Sample Diluent||1 vial, 20 mL||2-8°C, 6 months|
|Biotinylated Detection Ab Diluent||1 vial, 14 mL|
|HRP Conjugate Diluent||1 vial, 14 mL|
|Concentrated Wash Buffer (25×)||1 vial, 30 mL|
|Substrate Reagent||1 vial, 10 mL||2-8℃(Protect from light)|
|Stop Solution||1 vial, 10 mL||2-8°C|
|Plate Sealer||5 pieces|
|Certificate of Analysis||1 copy|
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human SEMA4D were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human SEMA4D were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
The recovery of Human SEMA4D spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
|Sample Type||Range (%)||Average Recovery (%)|
|EDTA plasma (n=8)||84-98||91|
|Cell culture media (n=8)||94-105||99|
Samples were spiked with high concentrations of Human SEMA4D and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C
2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C
3. Aspirate and wash the plate for 3 times
4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
6. Add 50μL Stop Solution
7. Read the plate at 450nm immediately. Calculation of the results
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