Human NFKB-p65 (Nuclear factor NF-kappa-B p65 subunit) ELISA Kit

Mfr.No.

E-EL-H1388

Sensitivity

0.1 ng/mL

Detection Range

0.16-10 ng/mL

Reactivity

Human

Assay Duration

3.5h

Assay Type

Sandwich

Description

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 Human NFKB-p65. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human NFKB-p65 and 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 Human NFKB-p65, 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 ± 2 nm. The OD value is proportional to the concentration of Human NFKB-p65. You can calculate the concentration of Human NFKB-p65 in the samples by comparing the OD of the samples to the standard curve.

Alternative Names

NFKB3 ; p65 ; RelA

Detection Method

Colormetric

Kit Components

An unopened kit can be stored at 2-8°C for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro ELISA Plate(Dismountable)	96T: 8 wells x12 strips 48T: 8 wells x6 strips	-20°C, 12 months
Reference Standard	96T: 2 vials 48T: 1 vial
Concentrated Biotinylated Detection Ab (100x)	96T: 1 vial, 120 uL 48T: 1 vial, 60 uL
Concentrated HRP Conjugate (100x)	96T: 1 vial, 120 uL 48T: 1 vial, 60 uL	-20°C(Protect from light), 12 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 12 months
Biotinylated Detection Ab Diluent	1 vial, 14 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25x)	1 vial, 30 mL
Substrate Reagent	1 vial, 10 mL	2-8°C(Protect from light)
Stop Solution	1 vial, 10 mL	2-8°C
Plate Sealer	5 pieces
Manual	1 copy
Certificate of Analysis	1 copy

Recovery

80%-120%. The recovery of Human NFKB-p65 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)	90-104	95
EDTA plasma (n=8)	92-105	100
Cell culture media (n=8)	86-97	92

Linearity

Samples were spiked with high concentrations of Human NF-κBp65 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 (%)	91-103	86-96	93-105
1:2	Average (%)	98	91	99
1:4	Range (%)	88-103	86-100	83-95
1:4	Average (%)	95	91	88
1:8	Range (%)	91-105	81-96	85-97
1:8	Average (%)	99	88	92
1:16	Range (%)	88-103	85-97	83-98
1:16	Average (%)	95	91	90

Specificity

This kit recognizes Human NFKB-p65 in samples. No significant cross-reactivity or interference between Human NFKB-p65 and analogues was observed.

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

Precision

Both intra-CV and inter-CV are < 10%.
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human NFKB-p65 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human NFKB-p65 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 (ng/mL)	0.54	1.49	3.97	0.52	1.39	3.93
Standard deviation	0.03	0.06	0.21	0.03	0.07	0.13
CV (%)	5.56	4.03	5.29	5.77	5.04	3.31

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 PDE4D .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 PDE4D 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 PDE4D ,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 PDE4D . The concentration of Human PDE4D in the samples can be calculated by comparing the OD of the samples to the standard curve.

Usage Notes

This ELISA kit applies to the in vitro quantitative determination of Human NFKB-p65 concentrations in serum, plasma and other biological fluids.

UniProt ID	Q04206

Applications	ELISA
Applications Notes	This ELISA kit applies to the in vitro quantitative determination of Human NFKB-p65 concentrations in serum, plasma and other biological fluids.
Research Area	Cancer, Cell Biology, Cardiovascular, Epigenetics and Nuclear Signaling, Metabolism, Microbiology, Neuroscience, Signal transduction

Storage	2-8°C/-20°C,12 months
Shipping	Ice bag

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SKU	Supplier No.	Size	Your price	Quantity
BHE10303814	E-EL-H1388	96T	$609		Add to Cart
BHE10303814	E-EL-H1388	48T	$487		Add to Cart

Human NFKB-p65 (Nuclear factor NF-kappa-B p65 subunit) ELISA Kit

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