Human sTNFRSF12A(Soluble Tumor Necrosis Factor Receptor Superfamily, Member 12A) ELISA Kit

Mfr.No.

E-EL-H2498

Antigen

sTNFRSF12A

Background

This kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human IgE .Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human IgE 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 Human IgE ,biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The OD value is measured spectrophotometrically at a wavelength of 450nm. The OD value is positively associated with the concentration of Human IgE . The concentration of Human IgE in the samples can be calculated by comparing the OD of the samples to the standard curve.

Description

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(pg/mL)	O.D	Average	Corrected
5000	2.439 2.471	2.455	2.385
2500	1.768 1.786	1.777	1.707
1250	1.014 1.004	1.009	0.9389999999999999
625	0.546 0.554	0.55	0.48
312.5	0.297 0.271	0.284	0.214
156.25	0.193 0.167	0.18	0.11
78.13	0.118 0.134	0.126	0.056
0	0.062 0.078	0.07000000000000001	--

Specificity

This kit recognizes Human sTNFRSF12A in samples. No significant cross-reactivity or interference between Human sTNFRSF12A 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 ELISA 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	1 vial, 10 mL	4℃(shading light)
Stop Solution	1 vial, 10 mL	4℃
Plate Sealer	5 pieces
Product Description	1 copy
Certificate of Analysis	1 copy

Detection Method

Colormetric

Detection Range

78.125--5000pg/mL

Linearity

Samples were spiked with high concentrations of Human sTNFRSF12A 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 (%)	89-103	87-101	87-101
1:2	Average (%)	95	94	92
1:4	Range (%)	94-109	84-95	85-97
1:4	Average (%)	100	90	91
1:8	Range (%)	91-103	86-99	87-101
1:8	Average (%)	97	91	93
1:16	Range (%)	91-105	80-93	87-103
1:16	Average (%)	96	86	94

Method Type

Sandwich

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human sTNFRSF12A were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human sTNFRSF12A 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)	263.00	772.30	1906.53	265.13	726.65	2065.20
Standard	deviation	17.33	33.60	65.97	16.76	40.11	109.04
C V (%)	6.59	4.35	3.46	6.32	5.52	5.28

Recovery

The recovery of Human sTNFRSF12A 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)	89-103	96
EDTA plasma (n=5)	92-103	98
Cell culture media (n=5)	87-100	92

Sample Type

Serum, Plasma, Cell supernatant

Sensitivity

46.88 pg/mL

Manual

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SKU	Supplier No.	Size	Your price
BHE10304677	E-EL-H2498	96T	$609	Add to Cart
BHE10304677	E-EL-H2498	96T*5	$2740.50	Add to Cart
BHE10304677	E-EL-H2498	96T*10	$5176.50	Add to Cart
BHE10304677	E-EL-H2498	48T	$487.20	Add to Cart

Human sTNFRSF12A(Soluble Tumor Necrosis Factor Receptor Superfamily, Member 12A) ELISA Kit

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