Human HMGB-1 (High Mobility Group Protein B1) CLIA Kit

Mfr.No.

E-CL-H0997

Sensitivity

7.5 pg/mL

Detection Range

12.5-800 pg/mL

Reactivity

Human

Assay Duration

3.5h

Assay Type

Sandwich

Description

This CLIA kit uses the Sandwich-CLIA principle. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Human HMGB-1. Standards or samples are added to the micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human HMGB-1 and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human HMGB-1, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Human HMGB-1. You can calculate the concentration of Human HMGB-1 in the samples by comparing the RLU value of the samples to the standard curve.

Alternative Names

HMGB1 ; HMG1 ; HMG3 ; SBP-1 ; Amphoterin

Detection Method

Chemiluminescence

Kit Components

An unopened kit can be stored at 2-8°C for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro CLIA Plate(Dismountable)	96T: 8 wells x12 strips 48T: 8 wells x6 strips	-20°C, 12 months
Reference Standard	96T: 2 vials 48T: 1 vial
Concentrated Biotinylated Detection Ab (100x)	96T: 1 vial, 120 uL 48T: 1 vial, 60 uL
Concentrated HRP Conjugate (100x)	96T: 1 vial, 120 uL 48T: 1 vial, 60 uL	-20°C(Protect from light), 12 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 12 months
Biotinylated Detection Ab Diluent	1 vial, 14 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25x)	1 vial, 30 mL
Substrate Reagent A	1 vial, 5 mL	2-8°C(Protect from light)
Substrate Reagent B	1 vial, 5 mL	2-8°C(Protect from light)
Plate Sealer	5 pieces
Manual	1 copy
Certificate of Analysis	1 copy

Recovery

80%-120%. The recovery of Human HMGB-1 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=8)	99-111	104
EDTA plasma(n=8)	94-111	102
Cell culture media(n=8)	86-99	91

Linearity

Samples were spiked with high concentrations of Human HMGB-1 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 (%)	93-109	103-118	103-120
1:2	Average (%)	101	109	109
1:4	Range (%)	95-111	94-107	85-98
1:4	Average (%)	103	99	90
1:8	Range (%)	93-109	86-100	101-114
1:8	Average (%)	99	93	108
1:16	Range (%)	88-101	85-99	88-101
1:16	Average (%)	95	91	94

Specificity

This kit recognizes Human HMGB-1 in samples. No significant cross-reactivity or interference between Human HMGB-1 and analogues was observed.

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

Precision

Both intra-CV and inter-CV are < 10%.
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human HMGB-1 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human HMGB-1 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)	38.94	71.20	397.79	37.00	72.83	379.26
Standard deviation	4.74	8.16	27.92	3.48	5.48	28.71
CV (%)	12.17	11.46	7.02	9.41	7.52	7.57

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

Usage Notes

This ELISA kit applies to the in vitro quantitative determination of Human HMGB-1 concentrations in serum, plasma and other biological fluids.

UniProt ID	P09429

Applications	CLIA
Applications Notes	This CLIA kit applies to the in vitro quantitative determination of Human HMGB-1 concentrations in serum, plasma and other biological fluids.
Research Area	Cancer, Epigenetics and Nuclear Signaling

Storage	2-8°C/-20°C,6 months
Shipping	Ice bag

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SKU	Supplier No.	Size	Your price	Quantity
BHE10300875	E-CL-H0997	96T	$682		Add to Cart
BHE10300875	E-CL-H0997	48T	$546		Add to Cart

Human HMGB-1 (High Mobility Group Protein B1) CLIA Kit

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