Human TNF-alpha(Tumor Necrosis Factor Alpha) CLIA Kit

Mfr.No.

E-CL-H0108

Antigen

TNF-α

Alternative Names

DIF ; TNF-alpha ; TNFA ; TNFSF2

Background

This kit uses Sandwich-CLIA as the method. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Human ANG1 Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human ANG1 and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain

Description

As the RLU 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(pg/mL)	RLU 1	RLU 2	Average RLU	Corrected RLU
500	49282	53792	51537	51508
250	19118	22878	20998	20969
125	9582	9210	9396	9367
62.5	4339	4685	4512	4483
31.25	2452	2146	2299	2270
15.625	1341	1159	1250	1221
7.81	735	745	740	711
0	29	29	29	--

Reactivity

Human

Specificity

This kit recognizes Human TNF-α in samples. No significant cross-reactivity or interference between Human TNF-α and analogues was observed.

Assay Time

4.5h

Components

An unopened kit can be stored at 4℃ for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro CLIA Plate(Dismountable)	8 wells ×12 strips	-20℃, 6 months
Reference Standard	2 vials
Concentrated Biotinylated Detection Ab (100×)	1 vial, 120 μL
Concentrated HRP Conjugate (100×)	1 vial, 120 μL	-20℃(shading light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	4℃, 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 A	1 vial, 5 mL	4℃ (shading light)
Substrate Reagent B	1 vial, 5 mL	4℃
Plate Sealer	5 pieces
Product Description	1 copy
Certificate of Analysis	1 copy

Detection Method

Chemiluminescence

Detection Range

7.813--500 pg/mL

Linearity

Samples were spiked with high concentrations of Human TNF-α 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 (%)	86-99	98-112	97-110
1:2	Average (%)	93	105	103
1:4	Range (%)	99-111	101-117	85-98
1:4	Average (%)	105	109	90
1:8	Range (%)	90-102	91-105	93-106
1:8	Average (%)	97	99	98
1:16	Range (%)	103-121	85-96	103-116
1:16	Average (%)	110	91	109

Method Type

Sandwich

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human TNF-α were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human TNF-α were tested on 3 different plates, 20 replicates in each plate.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (pg/mL)	25.60	58.03	169.01	27.58	54.45	158.50
Standard deviation	2.69	5.50	13.54	2.46	6.40	14.99
C V (%)	10.51	9.48	8.01	8.92	11.75	9.46

Recovery

The recovery of Human TNF-α 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=5)	100-113	107
EDTA plasma (n=5)	92-103	98
Cell culture media (n=5)	87-101	92

Sample Type

Serum, Plasma, Cell supernatant

Sensitivity

4.69 pg/mL

Uniprot	P01375

Manual

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SKU	Supplier No.	Size	Your price
BHE10300105	E-CL-H0108	96T	$682	Add to Cart
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BHE10300105	E-CL-H0108	96T*10	$5797	Add to Cart
BHE10300105	E-CL-H0108	48T	$545.60	Add to Cart

Human TNF-alpha(Tumor Necrosis Factor Alpha) CLIA Kit

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