Micro Fast ELISA Kits

MF-Mouse TNF-α(Tumor Necrosis Factor Alpha) ELISA Kit

Name: MF-Mouse TNF-α(Tumor Necrosis Factor Alpha) ELISA Kit
Brand: Reed Biotech
SKU: RE1060MF

Catalog #: RE1060MF

Manufacturer: Reed Biotech

Size: 96 T48 T

Price: on inquiry

Inquire / Order Manual (PDF)

Reactivity: Mouse
Sensitivity: 6.56 pg/mL
Assay Range: 10.94-700pg/mL
Sample Type: Serum, Plasma and Tissue homogenate and Other biological samples；

ELISA

Product Details

Product Name	MF-Mouse TNF-α(Tumor Necrosis Factor Alpha) ELISA Kit	Species	Mouse
Uniprot ID	P06804	Alternative Names	DIF, TNF-alpha, TNFA, TNFSF2, TNF-A, TNFα
Detection method	Sandwich	Sensitivity	6.56 pg/mL
Standard	700pg/mL	Detection Range	10.94-700pg/mL
Sample type	Serum, Plasma and Tissue homogenate and Other biological samples；Sample volume：25μL
Reaction time	3H	Research Area	Cardiovascular, Cell biology, Neuroscience, Cancer, Immunology, Metabolism
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 Mouse TNF-α. Samples (or Standards) are added to the micro ELISA plate wells and combined with the specific antibody and biotinylated detection antibody specific for Mouse TNF-α. Then Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Mouse TNF-α, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. 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 Mouse TNF-α. You can calculate the concentration of Mouse TNF-α 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.

(pg/mL)	OD	Corrected
700.00	2.080	1.995
350.00	1.207	1.123
175.00	0.589	0.505
87.50	0.397	0.313
43.75	0.219	0.135
21.88	0.156	0.072
10.94	0.119	0.035
0.00	0.084

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(pg/mL)	7.82	107.45	129.49	12.07	71.09	186.60
Standard deviation	0.70	6.03	7.61	0.66	4.02	7.33
C V (%)	3.21	6.62	3.48	2.84	5.93	3.27

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)	96-109	102
EDTA plasma (n=8)	81-95	88
Cell culture media (n=8)	85-97	91

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 (%)	86-97	95-102	91-102
1:2	Average (%)	95	101	97
1:4	Range (%)	83-96	95-102	89-99
1:4	Average (%)	84	97	92
1:8	Range (%)	91-103	98-106	88-104
1:8	Average (%)	103	101	102
1:16	Range (%)	96-105	87-98	84-98
1:16	Average (%)	100	90	98