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Rat ACTH(Adrenocorticotropic Hormone) ELISA Kit

  • Cat.No.:E-EL-R0048

  • Reactivity: Rat

To Purchase E-EL-R0048

  • 96T
  • 48T
  • 24T
  • 96T*5
  • 96T*10
Price: $495

Product Details


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.
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


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) 50.70 99.80 387.80 48.20 101.90 401.40
Standard deviation 3.10 5.30 15.90 3.10 5.20 14.50
CV (%) 6.11 5.31 4.10 6.43 5.10 3.61


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) 93-106 99
EDTA plasma(n=8) 91-107 98
Cell culture media(n=8) 86-100 93


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

Target Information

SynonymsCorticotropin, POMC, Adrenocorticotropin
Research AreaCancer, Metabolism, Neuroscience, Signal transduction

Assay Procedures

elisa assay procedure 1

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

elisa assay procedure 2

2. Aspirate and wash the plate for 3 times

elisa assay procedure 3

3. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times

elisa assay procedure 4

4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C

elisa assay procedure 5

5. Add 50μL Stop Solution

elisa assay procedure 6

6. Read the plate at 450nm immediately. Calculation of the results


    Acute exposure to microcystins affects hypothalamic-pituitary axes of male rats

    DOI: 10.1016/j.envpol.2022.120843

    Sample: serum
    Effects of acute exposure to microcystins on hypothalamic-pituitary-adrenal (HPA), -gonad (HPG) and -thyroid (HPT) axes of female rats

    DOI: 10.1016/j.scitotenv.2021.145196

    PMID: 34030373

    Sample: Serum
  3. 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: serum
  4. Pharmacological Reports (2021) IF: 3.027
    Development and treatment of cognitive inflexibility in sub-chronic stress–re-stress (SRS) model of PTSD

    DOI: 10.1007/s43440-020-00198-9

    Sample: Plasma
  5. Nutrition & Metabolism (2020) IF: 3.211
    The dysregulation of the hypothalamic–pituitary–adrenal axis in diet-induced prediabetic male Sprague Dawley rats

    DOI: 10.1186/s12986-020-00532-1

    Sample: plasma
  6. American Journal of Translational Research (2019) IF: 3.266
    Therapeutic effect of low-intensity pulsed ultrasound on temporomandibular joint injury induced by chronic sleep deprivation in rats

    PMID: 31312347

    Sample: serum
    Acupuncture exerts preventive effects in rats of chronic unpredictable mild stress: The involvement of inflammation in amygdala and brain-spleen axis

    DOI: 10.1016/j.bbrc.2023.01.046

    Sample: Serum,amygdala
    Activation of 5-HT1A Receptors in the Hypothalamic Paraventricular Nuclei Negatively Regulates Cytochrome P450 Expression and Activity in Rat Liver

    DOI: 10.1124/dmd.117.079632

    Sample: Serum,Tissue homogenate
  9. Experimental and Therapeutic Medicine (2018) IF: 2.447
    Analysis of gene expression and functional changes of adrenal gland in a rat model of kidney yang deficiency syndrome treated with Sini decoction

    DOI: 10.3892/etm.2018.6521

    PMID: 30214533

    Sample: Serum
    Lilium davidii extract alleviates p‑chlorophenylalanine‑induced insomnia in rats through modification of the hypothalamic-related neurotransmitters, melatonin and homeostasis of the hypothalamic-pituitary-adrenal axis

    DOI: 10.1080/13880209.2020.1812674

    PMID: 32924742

    Sample: serum
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