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Mouse ACTH(Adrenocorticotropic Hormone) ELISA Kit

Cat.No.:E-EL-M0079
Reactivity: Mouse

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96T
48T
24T
96T*5
96T*10

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Product Details

Properties

Assay type	Competitive-ELISA
Format	96T/48T
Assay time	2.0h
Detection range	15.63-1000 pg/mL
Sensitivity	9.38 pg/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 50μL
Specificity	This kit recognizes ACTH in samples. No significant cross-reactivity or interference between ACTH 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 ACTH concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with ACTH. During the reaction, ACTH in the sample or standard competes with a fixed amount of ACTH on the solid phase supporter for sites on the Biotinylated Detection Ab specific to ACTH. Excess conjugate and unbound sample or standard are washed away, and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns from blue to yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of ACTH in tested samples can be calculated 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℃
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 ACTH were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level ACTH 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)	52.20	119.30	479.60	47.00	128.20	475.90
Standard deviation	3.10	6.20	24.00	3.10	6.00	26.20
CV (%)	5.94	5.20	5.00	6.60	4.68	5.51

Recovery

The recovery of ACTH 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)	89-102	94
EDTA plasma(n=8)	88-104	95
Cell culture media(n=8)	89-102	96

Linearity

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

Target Information

Synonyms	Corticotropin, POMC, Adrenocorticotropin
Research Area	Cancer, Metabolism, Neuroscience, Signal transduction

Assay Procedures

	1. Add 50μL standard or sample to the wells, immediately add 50μL Biotinylated Detection Ab working solution to each well. Incubate for 45 min at 37°C
	2. Aspirate and wash the plate for 3 times
	3. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	5. Add 50μL Stop Solution
	6. Read the plate at 450nm immediately. Calculation of the results

Citations

CHINESE JOURNAL OF CHEMISTRY (2021) IF: 6
Uncarialins J‐M from Uncaria rhynchophylla and their anti‐depression mechanism in unpredictable chronic mild stress‐induced mice via activating 5‐HT1A receptor

DOI: 10.1002/cjoc.202000652

Sample: lung
FASEB JOURNAL (2021) IF: 5.192
High-fat diet-induced increases in glucocorticoids contribute to the development of non-alcoholic fatty liver disease in mice

DOI: 10.1096/fj.202101570R

PMID: 34959259
Sample: plasma
Journal of Functional Foods (2022) IF: 5.223
Juglanin ameliorates depression-like behavior in chronic unpredictable mild stress-induced mice by improving AMPK signaling

DOI: 10.1016/j.jff.2022.105263

Sample: serum
Food & Function (2021) IF: 5.396
Pediococcus acidilactici CCFM6432 mitigates chronic stress-induced anxiety and gut microbial abnormalities

DOI: 10.1039/D1FO01608C

PMID: 34704999
Sample: serum
PHYTOMEDICINE (2020) IF: 4.268
Uncaria rhynchophylla ameliorates unpredictable chronic mild stress-induced depression in mice via activating 5-HT1A receptor: Insights from transcriptomics

DOI: 10.1016/j.phymed.2020.153436

Sample: serum
Scientific Reports (2021) IF: 4.38
The role of inflammatory cytokines in anemia and gastrointestinal mucosal injury induced by foot electric stimulation

DOI: 10.1038/s41598-021-82604-7

PMID: 33542312
Sample: Serum,Tissue homogenate(hypothalamus,hypophysis )
BEHAVIOURAL BRAIN RESEARCH (2018) IF: 3.173
Antidepressant activity of vorinostat is associated with amelioration of oxidative stress and inflammation in a corticosterone-induced chronic stress model in mice

DOI: 10.1016/j.bbr.2018.02.009

PMID: 29452193
Sample: Serum
HORMONES AND BEHAVIOR (2017) IF: 3.378
What is stressful for females? Differential effects of unpredictable environmental or social stress in CD1 female mice

DOI: 10.1016/j.yhbeh.2017.11.013

Sample: plasma
Scientific Reports (2020) IF: 3.998
Long term transcriptional and behavioral effects in mice developmentally exposed to a mixture of endocrine disruptors associated with delayed human neurodevelopment

DOI: 10.1038/s41598-020-66379-x

PMID: 32518293
Sample: Serum
JOURNAL OF PHYSIOLOGY AND PHARMACOLOGY (2018) IF: 2.478
High activity of the endogenous opioid system and acute but not chronic stress influence experimental colitis development in mice

DOI: 10.26402/jpp.2018.5.11

Sample: plasma
RESEARCH IN VETERINARY SCIENCE (2019) IF: 1.751
Amelioration effects of Kaempferol on immune response following chronic intermittent cold-stress

DOI: 10.1016/j.rvsc.2019.08.012

PMID: 31412308
Sample: Serum,Tissue homogenate
Sleep and Biological Rhythms (2018) IF: 0.655
Evaluation of behavior and endocrinological changes after REM sleep deprivation-induced mania-like behavior in mice

DOI: 10.1007/s41105-018-0166-6

Sample: serum

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