Human PADI4(Peptidyl Arginine Deiminase Type IV) CLIA Kit

Mfr.No.

E-CL-H0866

Antigen

PADI4

Alternative Names

PAD ; PAD4 ; PADI5 ; PDI4 ; PDI5

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	55460	62702	59081	59046
2000	26160	29558	27859	27824
1000	14002	12966	13484	13449
500	6350	6860	6605	6570
250	3349	3137	3243	3208
125	1649	1513	1581	1546
62.50	714	796	755	720
0	33	37	35	--

Specificity

This kit recognizes Human PADI4 in samples. No significant cross-reactivity or interference between Human PADI4 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 PADI4 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 (%)	96-108	100-112	102-116
1:2	Average (%)	101	107	108
1:4	Range (%)	96-110	90-103	90-104
1:4	Average (%)	102	97	98
1:8	Range (%)	91-106	101-115	98-111
1:8	Average (%)	97	107	104
1:16	Range (%)	101-117	100-116	90-102
1:16	Average (%)	108	107	95

Method Type

Sandwich

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human PADI4 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human PADI4 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)	196.88	406.01	1941.91	213.15	388.35	1806.50
Standard deviation	21.26	39.75	205.84	27.41	35.34	134.04
C V (%)	10.80	9.79	10.60	12.86	9.10	7.42

Recovery

The recovery of Human PADI4 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)	98-110	103
EDTA plasma (n=5)	99-114	106
Cell culture media (n=5)	88-100	95

Sample Type

Serum, Plasma, Cell supernatant

Sensitivity

37.5 pg/mL

Uniprot	Q9UM07

Manual

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Human PADI4(Peptidyl Arginine Deiminase Type IV) CLIA Kit

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