Human 12-LO(Arachidonate 12-LipoXygenase) CLIA Kit

Mfr.No.

E-CL-H1438

Antigen

12-LO

Alternative Names

ALOX12 ; LOX12 ; 12-LOX ; 12LPG ; LOG12

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
4000	54424	63738	59081	59046
2000	26225	29493	27859	27824
1000	14621	12347	13484	13449
500	6433	6777	6605	6570
250	3320	3166	3243	3208
125	1629	1533	1581	1546
62.50	742	768	755	720
0	33	37	35	--

Reactivity

Human

Specificity

This kit recognizes Human 12-LO in samples. No significant cross-reactivity or interference between Human 12-LO 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

62.5--4000 pg/mL

Linearity

Samples were spiked with high concentrations of Human 12-LO 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 (%)	88-102	86-96	95-112
1:2	Average (%)	94	91	103
1:4	Range (%)	89-104	86-97	85-98
1:4	Average (%)	96	92	91
1:8	Range (%)	98-115	92-108	91-106
1:8	Average (%)	105	100	98
1:16	Range (%)	97-113	83-95	94-109
1:16	Average (%)	105	90	99

Method Type

Sandwich

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human 12-LO were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human 12-LO 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)	211.96	500.64	1517.01	213.47	452.44	1571.15
Standard deviation	26.11	54.32	134.26	26.15	50.31	160.57
C V (%)	12.32	10.85	8.85	12.25	11.12	10.22

Recovery

The recovery of Human 12-LO 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)	88-103	96
EDTA plasma (n=5)	87-100	93
Cell culture media (n=5)	95-108	102

Sample Type

Serum, Plasma, Cell supernatant

Sensitivity

37.5 pg/mL

Uniprot	P18054

Manual

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SKU	Supplier No.	Size	Your price
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Human 12-LO(Arachidonate 12-Lipoxygenase) CLIA Kit

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