Human IL-12 p40(Interleukin 12 p40) CLIA Kit

Mfr.No.

E-CL-H0143

Antigen

IL-12p40

Alternative Names

IL12B ; IL-12B ; CLMF2 ; NKSF2 ; p40 ; IL-12/IL-23 p40

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
800	48002	57282	52642	52616
400	20683	22785	21734	21708
200	9862	9554	9708	9682
100	4404	4700	4552	4526
50	2328	2048	2188	2162
25	1090	1030	1060	1034
12.50	492	526	509	483
0	25	27	26	--

Specificity

This kit recognizes Human IL-12p40 in samples. No significant cross-reactivity or interference between Human IL-12p40 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

12.5--800 pg/mL

Linearity

Samples were spiked with high concentrations of Human IL-12p40 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 (%)	84-98	101-120	100-116
1:2	Average (%)	91	109	108
1:4	Range (%)	101-118	86-101	87-99
1:4	Average (%)	108	93	93
1:8	Range (%)	93-107	103-117	90-105
1:8	Average (%)	98	109	97
1:16	Range (%)	103-120	94-106	103-117
1:16	Average (%)	110	100	109

Method Type

Sandwich

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human IL-12p40 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human IL-12p40 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)	39.18	69.10	293.76	40.45	72.22	264.60
Standard deviation	3.32	6.85	26.97	4.62	8.14	24.08
C V (%)	8.47	9.91	9.18	11.42	11.27	9.10

Recovery

The recovery of Human IL-12p40 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	95
EDTA plasma (n=5)	92-106	97
Cell culture media (n=5)	100-111	106

Sample Type

Serum, Plasma, Cell supernatant

Sensitivity

7.5 pg/mL

Uniprot	P29460

Manual

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SKU	Supplier No.	Size	Your price
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Human IL-12 p40(Interleukin 12 p40) CLIA Kit

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