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Human ACTH(Adrenocorticotropic Hormone) ELISA Kit

Cat.No.:E-EL-H0137
Reactivity: Human

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96T*5
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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)	45.70	130.90	461.60	45.10	124.70	448.60
Standard deviation	3.20	6.80	14.30	2.30	6.40	19.70
CV (%)	7.00	5.19	3.10	5.10	5.13	4.39

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)	91-104	98
EDTA plasma(n=8)	85-100	91
Cell culture media(n=8)	86-98	93

Linearity

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

Target Information

Synonyms	Corticotropin, 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

ENVIRONMENTAL POLLUTION (2022) IF: 9.988
Neuroendocrine stress hormones associated with short-term exposure to nitrogen dioxide and fine particulate matter in individuals with and without chronic obstructive pulmonary disease: A panel study in Beijing, China

DOI: 10.1016/j.envpol.2022.119822

PMID: 35870527
Sample: serum
Computational and Structural Biotechnology Journal (2020) IF: 6.018
Lipid metabolic signatures deviate in sepsis survivors compared to non-survivors

DOI: 10.1016/j.csbj.2020.11.009

PMID: 33304464
Sample: blood
Nutrients (2021) IF: 5.719
Differential Health Effects on Inflammatory, Immunological and Stress Parameters in Professional Soccer Players and Sedentary Individuals after Consuming a Synbiotic. A Triple-Blinded, Randomized, Placebo-Controlled Pilot Study

DOI: 10.3390/nu13041321

PMID: 33923663
Sample: Whole Blood

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Reviews/Q&A

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Reviews
Q&A

I****rSubmitted [ Aug 20 2018 ]

Asked: Good Morning !! I need to know if this kit (E-EL-H0137) can be used (validated) with saliva samples. So if you can send the options to me I appreciate it. Thank you

adminSubmitted [ Aug 20 2018 ]

Answered: Thank you Itamar for your question. For this kit with Cat. #E-EL-H0137, it can be used with saliva samples. Please feel free to contact us if any other question. Thank you.

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