Our products are for in vitro research use only, not for clinical diagnosis

Nickname | quit

Recently added:

in total 0 items Total 0

Language:

Home
Product
Research
Services
News
About

All

All
Products
News
Introduction
FAQ
Video
Gallery

All

All
Products
News
Introduction
FAQ
Video
Gallery

Home page

Collection

Contrast

No .

E-EL-H1554c

Human HMGB-1(Highmobility group protein B1) ELISA Kit

Item No:

E-EL-H1554c

Chinese Name:

Alias:

HMGB1, HMG1, HMG3, SBP-1, Amphoterin

Keywords:

0px

Classification:

Recombinant protein

Retail Price

¥ 3000

Market Price

¥ 1

This kit is used for the in vitro quantification of HMGB-1 in human serum, plasma or other biological fluids.

Spec
- 48T
- 96T
- 96T*5

Quantity

Inventory surplus

隐藏域元素占位

Product Comparison

Total X Clear all

Describe

Use

　　This kit is used for the in vitro quantification of HMGB-1 in human serum, plasma or other biological fluids.

Principle of detection

　　This kit adopts double antibody sandwich ELISA method. Anti-Human HMGB-1 antibody is used to coat the ELISA plate. Human HMGB-1 in the sample or standard will bind to the coating antibody during the experiment, and the free components are washed away. Biotinylated anti-Human HMGB-1 antibody and horseradish peroxidase-labeled affinity are added sequentially. The anti-human HMGB-1 antibody binds to the human HMGB-1 bound to the encapsulated antibody, and the biotin binds specifically to the affin to form an immune complex, and the free components are washed away. A chromogenic substrate (TMB) is added, and TMB appears blue catalyzed by horseradish peroxidase and turns yellow after the addition of termination solution. The concentration of HMGB-1 in the sample was calculated by plotting the standard curve.

Reaction Type	Sandwich method
Specification	96T
Reaction time	3.5h
Reactivity	Human
Detection method	Colormetric
Detection range	31.25—2000 pg/mL
Sensitivity	18.75 pg/mL
Sample volume	100μL
Sample type	Serum, plasma or other biological fluids

Specificity

　　Detects human HMGB-1 in samples with no significant cross-reactivity with other related proteins.

Repeatability

　　Intra-plate and inter-plate coefficients of variation <10%.

Typical data

　 The OD values of the standard curves may vary due to different operating conditions (e.g. operator, pipetting technique, plate washing technique and stabilization conditions). The following data and curves are for reference only. Experimenters need to establish standard curves according to their own experiments.

(pg/mL)	O.D	Average	Corrected
2000	2.443 2.447	2.445	2.374
1000	1.718 1.764	1.741	1.67
500	0.988 0.984	0.986	0.915
250	0.466 0.498	0.482	0.411
125	0.265 0.247	0.256	0.185
62.5	0.187 0.161	0.174	0.103
31.25	0.118 0.13	0.124	0.053
0	0.069 0.073	0.070	--

Precision　

　　Intra-plate precision: 20 times on 1 plate for low, medium and high concentration samples.
　　Intra-plate precision: 20 times on 3 plates for low, medium and high concentrations.

Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (pg/mL)	99.32	202.75	693.26	108.96	208.03	746.59
Standard deviation	5.84	8.62	35.56	5.86	9.76	26.06
C V (%)	5.88	4.25	5.13	5.38	4.69	3.49

Recovery

　　Recovery experiments were performed by adding known concentrations of target proteins to five different samples, and the recovery range and average recovery were obtained.

Sample Type	Range (%)	Average Recovery (%)
Serum (n=8)	94-108	100
EDTA plasma (n=8)	94-107	100
Cell culture media (n=8)	93-105	99

Linearity

　　Recovery experiments were performed by adding known concentrations of target proteins to 5 samples, and the range of recovery rates and average recovery rates were obtained. Five samples were diluted 2, 4, 8, and 16 times for recovery experiments, and the range of recovery rates and average recovery rates were obtained.

	Serum (n=5)	EDTA plasma (n=5)	Cell culture media (n=5)
1:2	Range (%)	98-111	92-105	85-100
1:2	Average (%)	105	98	92
1:4	Range (%)	90-103	84-100	88-102
1:4	Average (%)	98	91	94
1:8	Range (%)	87-99	83-93	85-96
1:8	Average (%)	94	88	90
1:16	Range (%)	88-103	85-96	86-99
1:16	Average (%)	94	90	91

Kit composition and storage

　　Unopened kits can be stored at 2-8°C for one week; if the kit is used after one week, please unpack the kit and store each component separately according to the conditions in the table below.

Chinese name	English Name	Specification	Storage conditions
ELISA enzyme standard plate (detachable)	Micro ELISA Plate(Dismountable)	8 holes×12 strips	-20℃, can be stored for 6 months
Lyophilized standards	Reference Standard	2 sticks
Concentrated biotinylated antibody(100×)	Concentrated Biotinylated Detection Ab	1 x 120 μL
Concentrated HRP enzyme conjugate(100×)	Concentrated HRP Conjugate	1 x 120 μL	-20℃(keep away from light), can be stored for 6 months
Standard & Sample Diluent	Reference Standard & Sample Diluent	1 bottle of 20 mL	2-8℃. Can be stored for 6 months
Biotinylated antibody diluent	Biotinylated Detection Ab Diluent	1 bottle 14 mL
Enzyme Conjugate Diluent	HRP Conjugate Diluent	1 bottle 14 mL
Concentrated Washing Solution(25×)	Concentrated Wash Buffer (25×)	1 bottle 30 mL
Substrate solution (TMB)	Substrate Reagent	1 bottle 10 mL	2-8℃(keep away from light)
Reaction termination solution	Stop Solution	1 bottle 10 mL	2-8℃
Plate lamination	Plate Sealer	5 sheets
Product manual	Product Description	1 copy
Quality inspection report	Certificate of Analysis	1 copy

　　Note: All reagent bottles must be tightly capped to prevent evaporation and microbial contamination.
　　The volume of reagents is subject to the actual shipping instructions. The relevant reagents will be slightly more than the volume indicated on the label when dispensing, please measure rather than pour directly when using.

Items required for the test

　　1. enzyme calibrator (450nm wavelength filter)
　　2. High precision pipettes, EP tubes and disposable tips: 0.5-10 μL, 2-20 μL, 20-200 μL, 200-1000 μL
　　3. 37°C thermostat, double-distilled water or deionized water
　　4. absorbent paper

Operation steps

	1. Immediately after adding 50 μL of standard working solution or sample to the corresponding plate wells, add 50 μL of biotinylated antibody working solution to each well and incubate at 37°C for 45 minutes.
	2. Discard the liquid in the plate and wash the plate 3 times
	3. Add 100μL of HRP enzyme conjugate working solution to each well, incubate at 37℃ for 30 minutes, discard the liquid in the plate, and wash the plate 5 times
	4. Add 90 μL of substrate solution to each well and incubate at 37°C for 15 minutes.
	5. Add 50μL of termination solution to each well
	6. Read immediately at 450 nm and process the data