Our Elisa Kit products are perfect for the analysis of biological fluids, including Serum, plasma, tissue homogenates, cell lysates, cell culture supernatants, and other biological fluids.
This assay has high sensitivity and excellent specificity for the detection of Tumor Necrosis Factor Alpha (TNFa). No significant cross-reactivity or interference between Tumor Necrosis Factor Alpha (TNFa) and analogs was observed.
Recovery
Matrices listed below were spiked with a certain level of recombinant Tumor Necrosis Factor Alpha (TNFa). The recovery rates were calculated by comparing the measured value to the expected amount of Tumor Necrosis Factor Alpha (TNFa) in samples.
Matrix
Recovery range (%)
Average(%)
serum(n=5)
83-99
94
EDTA plasma(n=5)
81-92
88
heparin plasma(n=5)
82-98
88
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle, and high-level Tumor Necrosis Factor Alpha (TNFa) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle, and high-level Tumor Necrosis Factor Alpha (TNFa) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CV<10% Inter-assay: CV<12%Â
Linearity
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Tumor Necrosis Factor Alpha (TNFa) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample
1:2
1:4
1:8
1:16
serum(n=5)
80-98%
94-104%
95-104%
79-101%
EDTA plasma(n=5)
84-95%
81-102%
84-94%
78-103%
heparin plasma(n=5)
95-102%
98-105%
82-101%
86-95%
Stability
The loss rate of activity determines the stability of the kit. The loss rate of this kit is less than 5% within the expiration date under appropriate storage conditions. To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, and incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.
Packaging & Shipping
Reagents and materials provided
Reagents
Quantity
Reagents
Quantity
Pre-coated, ready-to-use 96-well strip plate
1
Plate sealer for 96 wells
4
Standard
2
Standard Diluent
1×20mL
Detection Reagent A
1×120µL
Assay Diluent A
1×12mL
Detection Reagent B
1×120µL
Assay Diluent B
1×12mL
TMB Substrate
1×9mL
Stop Solution
1×6mL
Wash Buffer (30 × concentrates)
1×20mL
Instruction manual
1
Â
Procedure
1. Prepare all reagents, samples, and standards; 2. Add 100µL standard or sample to each well. Incubate for 1 hour at 37°C; 3. Aspirate and add 100µL prepared Detection Reagent A. Incubate for 1 hour at 37°C; 4. Aspirate and wash 3 times; 5. Add 100µL prepared Detection Reagent B. Incubate for 30 minutes at 37°C; 6. Aspirate and wash 5 times; 7. Add 90µL Substrate Solution. Incubate for 10-20 minutes at 37°C; 8. Add 50µL Stop Solution. Read at 450nm immediately.Â
Additional information
Brand
Cloud Clone
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