Human NFE2L2(Nuclear Factor, Erythroid DerIVed 2, Like 2) CLIA Kit

Mfr.No.

E-CL-H1005

Antigen

NFE2L2

Alternative Names

NRF2 ; NF-E2-Related Factor 2

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
2000	55042	56400	55721	55693
1000	22312	25636	23974	23946
500	11918	10146	11032	11004
250	4903	5685	5294	5266
125	2755	2461	2608	2580
62.5	1404	1218	1311	1283
31.25	628	720	674	646
0	27	29	28	--

Specificity

This kit recognizes Human NFE2L2 in samples. No significant cross-reactivity or interference between Human NFE2L2 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

31.25--2000 pg/mL

Linearity

Samples were spiked with high concentrations of Human NFE2L2 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 (%)	102-117	94-106	93-106
1:2	Average (%)	108	100	100
1:4	Range (%)	103-115	102-119	100-118
1:4	Average (%)	109	108	108
1:8	Range (%)	93-105	95-110	100-114
1:8	Average (%)	99	101	106
1:16	Range (%)	85-98	93-110	95-106
1:16	Average (%)	90	100	100

Method Type

Sandwich

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human NFE2L2 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human NFE2L2 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)	109.03	295.14	683.64	100.62	322.59	743.07
Standard deviation	11.37	33.29	63.99	10.90	28.36	66.65
C V (%)	10.43	11.28	9.36	10.83	8.79	8.97

Recovery

The recovery of Human NFE2L2 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)	98-111	105
EDTA plasma (n=5)	100-114	107
Cell culture media (n=5)	86-98	93

Sample Type

Serum, Plasma, Cell supernatant

Sensitivity

18.75 pg/mL

Uniprot	Q16236

Manual

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Human NFE2L2(Nuclear Factor, Erythroid Derived 2, Like 2) CLIA Kit

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