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Mouse Hepc(Hepcidin) ELISA Kit

Citations(12)Uniprot : Q9EQ21

Cat.No.:E-EL-M0671
Reactivity: Mouse

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96T
48T
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96T*5
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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	125.00-8000 pg/mL
Sensitivity	75.00 pg/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Mouse Hepc in samples. No significant cross-reactivity or interference between Mouse Hepc and analogues was observed.
Reproducibility	Both intra-CV and inter-CV are < 10%.
Application	This ELISA kit applies to the in vitro quantitative determination of Mouse Hepc concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Mouse Hepc. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Mouse Hepc 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 Mouse Hepc, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Mouse Hepc. You can calculate the concentration of Mouse Hepc in the samples by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ 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 ELISA Plate(Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	-20℃, 6 months
Reference Standard	96T: 2 vials 48T: 1 vial
Concentrated Biotinylated Detection Ab (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL	-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 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	1 vial, 10 mL	2-8℃(Protect from light)
Stop Solution	1 vial, 10 mL	2-8°C
Plate Sealer	5 pieces
Manual	1 copy
Certificate of Analysis	1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse Hepc were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse Hepc 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)	385.00	686.40	3381.60	352.70	696.90	3600.60
Standard deviation	23.50	30.20	121.70	24.70	38.30	144.00
CV (%)	6.10	4.40	3.60	7.00	5.50	4.00

Recovery

The recovery of Mouse Hepc 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)	87-101	94
EDTA plasma (n=8)	89-103	97
Cell culture media (n=8)	89-102	94

Linearity

Samples were spiked with high concentrations of Mouse Hepc and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database Links	SwissProt: Q9EQ21
Synonyms	Hepcidin Antimicrobial Peptide
Research Area	Cancer, Cardiovascular, Metabolism, Signal transduction

Assay Procedures

	1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C
	2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C
	3. Aspirate and wash the plate for 3 times
	4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	6. Add 50μL Stop Solution
	7. Read the plate at 450nm immediately. Calculation of the results

Citations

FOOD AND CHEMICAL TOXICOLOGY (2021) IF: 6.025
Perinatal and early-life cobalt exposure impairs essential metal metabolism in immature ICR mice

DOI: 10.1016/j.fct.2021.111973

PMID: 33421458
Sample: Serum
Antioxidants (2021) IF: 6.313
Epigallocatechin-3-Gallate Suppresses BMP-6-Mediated SMAD1/5/8 Transactivation of Hepcidin Gene by Inducing SMILE in Hepatocytes

DOI: 10.3390/antiox10101590

PMID: 34679725
Sample: AML12 cell,
Antioxidants (2020) IF: 5.014
Epigallocatechin-3-Gallate (EGCG)-Inducible SMILE Inhibits STAT3-Mediated Hepcidin Gene Expression

DOI: 10.3390/antiox9060514

PMID: 32545266
Sample: Serum,Cell culture medium
Frontiers in Immunology (2018) IF: 5.511
Ferritin H Deficiency in Myeloid Compartments Dysregulates Host Energy Metabolism and Increases Susceptibility to Mycobacterium tuberculosis Infection

DOI: 10.3389/fimmu.2018.00860

Sample: serum
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES (2022) IF: 5.924
Melatonin Regulates Iron Homeostasis by Inducing Hepcidin Expression in Hepatocytes

DOI: 10.3390/ijms23073593

PMID: 35408955
Sample: serum,AML12 cell
Nutrients (2020) IF: 4.546
Omega-3 Fatty Acid and Iron Supplementation Alone, but Not in Combination, Lower Inflammation and Anemia of Infection in Mycobacterium tuberculosis-Infected Mice

DOI: 10.3390/nu12092897

PMID: 32971969
Sample: Plasma
Frontiers in Physiology (2022) IF: 4.566
A Reproducible Mouse Model of Moderate CKD With Early Manifestations of Osteoblastic Transition of Cardiovascular System

DOI: 10.3389/fphys.2022.897179

PMID: 35574469
Sample: serum
Frontiers in Immunology (2019) IF: 4.716
Ferritin Light Chain Confers Protection Against Sepsis-Induced Inflammation and Organ Injury

DOI: 10.3389/fimmu.2019.00131

PMID: 30804939
Sample: Serum
JOURNAL OF MOLECULAR ENDOCRINOLOGY (2018) IF: 3.297
Hepcidin is an endogenous protective factor for osteoporosis by reducing iron levels

DOI: 10.1530/JME-17-0301

Sample: Serum
JOURNAL OF TRACE ELEMENTS IN MEDICINE AND BIOLOGY (2021) IF: 3.849
Calcitonin increases hepatic hepcidin expression through the BMP6 of kidney in mice

DOI: 10.1016/j.jtemb.2021.126796

PMID: 34098241
Sample: serum
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS (2019) IF: 2.705
Increased hepcidin in hemorrhagic plaques correlates with iron-stimulated IL-6/STAT3 pathway activation in macrophages

DOI: 10.1016/j.bbrc.2019.05.123

Sample: mouse peritoneal macrophages (MPMs)
JOURNAL OF CELLULAR BIOCHEMISTRY (2018) IF: 2.959
The α1‐adrenergic receptor is involved in hepcidin upregulation induced by adrenaline and norepinephrine via the STAT3 pathway

DOI: 10.1002/jcb.26715

Sample: serum

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