Human UBA52(Ubiquitin A 52 Residue Ribosomal Protein Fusion Product 1) CLIA Kit

Mfr.No.

E-CL-H1378

Antigen

UBA52

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	53212	64950	59081	59046
2000	27523	28195	27859	27824
1000	13700	13268	13484	13449
500	6434	6776	6605	6570
250	3361	3125	3243	3208
125	1646	1516	1581	1546
62.50	733	777	755	720
0	34	36	35	--

Specificity

This kit recognizes Human UBA52 in samples. No significant cross-reactivity or interference between Human UBA52 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 UBA52 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 (%)	95-107	101-113	90-103
1:2	Average (%)	102	107	96
1:4	Range (%)	96-110	88-98	96-109
1:4	Average (%)	103	93	104
1:8	Range (%)	98-114	93-106	88-103
1:8	Average (%)	105	99	96
1:16	Range (%)	97-113	92-104	89-99
1:16	Average (%)	104	99	94

Method Type

Sandwich

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human UBA52 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human UBA52 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)	206.43	617.17	1877.26	190.28	606.66	2035.73
Standard deviation	24.07	61.41	121.08	20.30	69.64	216.40
C V (%)	11.66	9.95	6.45	10.67	11.48	10.63

Recovery

The recovery of Human UBA52 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)	96-109	104
EDTA plasma (n=5)	99-116	106
Cell culture media (n=5)	97-112	103

Sample Type

Serum, Plasma, Cell supernatant

Sensitivity

37.5 pg/mL

Uniprot	P62987

Manual

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Human UBA52(Ubiquitin A 52 Residue Ribosomal Protein Fusion Product 1) CLIA Kit

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