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Endocrinology

ACTH ELISA

Catalog Number: 
M046006
Qty/Size: 
96 wells

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Adrenocorticotropic hormone (ACTH) ELISA kit to measure ACTH levels in human, mouse and rat plasma samples.

Overview

Overview: 

ACTH (Adrenocorticotropic hormone) or corticotropin is a 39-amino acid peptide hormone (MW=4500) secreted by the pituitary to regulate the production of steroid hormones by the adrenal cortex. ACTH secretion from the anterior pituitary is controlled by both a classical negative feedback control mechanism and CNS-stress mediated control system. Various types of stress or pain perceived in higher levels of the brain modulate secretion of the hypothalamic neurosecretory hormone, corticotropin releasing hormone (CRH), a 41-amino acid peptide. CRH stimulates pituitary ACTH secretion. The second peptide that modulates ACTH secretion is vasopressin (AVP). AVP secretion is also stimulated by stress and acts synergistically with CRH to increase ACTH secretion in the pituitary portal circulation. ACTH increases the synthesis and release of all adrenal sterioids, aldosterone, cortisol and adrenal androgens. It is the principal modulator of cortisol, the most important glucocorticoid in man. As the cortisol level in blood increases, release of ACTH is inhibited directly at the pituitary level. Through this same mechanism, decreasing cortisol levels lead to elevated ACTH levels.

 

Biologically active ACTH results from enzymatic cleavage of a large precursor molecule, pro-opiomelanocortin (POMC). This molecule contains within its structure the amino acid sequences of ACTH, Pro-ACTH, -melanocyte stimulating hormone, lipotropin, as well as endorphin and the enkephalins. Because the reaction in immunoassays is determined by antigenic structure, not biological function, the usual ACTH RIA reacts with POMC, Pro-ACTH, ACTH and some fragments of the ACTH.

Data/Specifications

Data/Specifications: 

Species: human, mouse, rat

Sample Type:  plasma

Sample Size: 200 uL

Standard Curve Range:  0 - 500 pg/mL

Sensitivity:  0.46 pg/mL

Assay Length:   4.5 hours

 

ACTH may also be referred to by the following terms:

ACTH ELISA kit, Adrenocorticotropic Hormone ELISA kit, Adrenocorticotropin ELISA kit, Alpha-Melanocyte-Stimulating Hormone ELISA kit, Alpha-MSH ELISA kit, Beta-Endorphin ELISA kit, Beta-LPH ELISA kit, Beta-Melanocyte-Stimulating Hormone ELISA kit, Beta-MSH ELISA kit, CLIP ELISA kit, Corticotropin-like Intermediary Peptide ELISA kit, Corticotropin-Lipotropin ELISA kit, Corticotropin-Lipotropin ELISA kit, Gamma-LPH ELISA kit, Gamma-MSH ELISA kit, Lipotropin Beta ELISA kit, Lipotropin Gamma ELISA kit, LPH ELISA kit, Melanotropin Alpha ELISA kit, Melanotropin Beta ELISA kit, Melanotropin Gamma ELISA kit, Met-Enkephalin ELISA kit, MSH ELISA kit, NPP ELISA kit, POC ELISA kit, Pro-ACTH-endorphin ELISA kit, Pro-OpioMelanocortin ELISA kit, ProopioMelanocortin ELISA kit, ProopioMelanocortin PreProProtein ELISA kit

Literature/Support

Literature/Support: 

Product Insert:

ACTH ELISA Insert (PDF)

 

Articles/FAQ:

Stress and Autoimmune Disease: the role of ACTH (blog post)

ELISA Data Reduction Guide

 

 

Rasmusson, A. M., King, M. W., Valovski, I., Gregor, K., Scioli-Salter, E., Pineles, S. L., ... & Pinna, G. (2019). Relationships between cerebrospinal fluid GABAergic neurosteroid levels and symptom severity in men with PTSD. Psychoneuroendocrinology, 102, 95-104.

 

Rasmusson, A. M., King, M. W., Valovski, I., Gregor, K., Scioli-Salter, E., Pineles, S. L., ... & Pinna, G. (2019). Relationships between cerebrospinal fluid GABAergic neurosteroid levels and symptom severity in men with PTSD. Psychoneuroendocrinology, 102, 95-104.

 

Basham, K. J., Rodriguez, S., Turcu, A. F., Lerario, A. M., Logan, C. Y., Rysztak, M. R., ... & Nusse, R. (2019). A ZNRF3-dependent Wnt/β-catenin signaling gradient is required for adrenal homeostasis. Genes & development33(3-4), 209-220.

 

Yasuda, A., Seki, T., Kametani, Y., Koizumi, M., Kitajima, N., Oki, M., ... & Fukagawa, M. (2019). Glucocorticoid Receptor Antagonist Administration Prevents Adrenal Gland Atrophy in an ACTH-Independent Cushing’s Syndrome Rat Model. International journal of endocrinology2019.

 

Asari, Y., Kageyama, K., Sugiyama, A., Kogawa, H., Niioka, K., & Daimon, M. (2019). Lapatinib decreases the ACTH production and proliferation of corticotroph tumor cells. Endocrine journal, EJ18-0491.

 

Ergang, P., Mikulecká, A., Vodicka, M., Vagnerová, K., Miksik, I., & Pacha, J. (2018). Social defeat stimulates local glucocorticoid regeneration in lymphoid organs. Endocrine connections1(aop).

 

Kameda, H., Yamamoto, M., Tone, Y., Tone, M., & Melmed, S. (2018). Proton Sensitivity of Corticotropin-Releasing Hormone Receptor 1 Signaling to Proopiomelanocortin in Male Mice. Endocrinology160(2), 276-291.

 

Takizawa, N., Tanaka, S., Oe, S., Koike, T., Yoshida, T., Hirahara, Y., ... & Yamada, H. (2018). Involvement of DHH and GLI1 in adrenocortical autograft regeneration in rats. Scientific reports8(1), 14542.

 

Lu, J., Montgomery, B. K., Chatain, G. P., Bugarini, A., Zhang, Q., Wang, X., ... & Chittiboina, P. (2018). Corticotropin releasing hormone can selectively stimulate glucose uptake in corticotropinoma via glucose transporter 1. Molecular and cellular endocrinology470, 105-114

 

Wang, D., Cai, M., Wang, T., Zhao, G., Huang, J., Wang, H., ... & Wang, Y. (2018). Theanine supplementation prevents liver injury and heat shock response by normalizing hypothalamic-pituitaryadrenal axis hyperactivity in mice subjected to whole body heat stress. Journal of Functional Foods45, 181-189.

 

Ziko, I., Sominsky, L., De Luca, S. N., Lelngei, F., & Spencer, S. J. (2018). Acylated ghrelin suppresses the cytokine response to lipopolysaccharide and does so independently of the hypothalamic-pituitary-adrenal axis. Brain, behavior, and immunity.

 

Lu, J., Chatain, G. P., Bugarini, A., Wang, X., Maric, D., Walbridge, S., ... & Chittiboina, P. (2017). Histone deacetylase inhibitor SAHA is a promising treatment of cushing disease. The Journal of Clinical Endocrinology & Metabolism102(8), 2825-2835.

 

Asari, Y., Kageyama, K., Nakada, Y., Tasso, M., Takayasu, S., Niioka, K., ... & Daimon, M. (2017). Inhibitory effects of a selective Jak2 inhibitor on adrenocorticotropic hormone production and proliferation of corticotroph tumor AtT20 cells. OncoTargets and therapy10, 4329.

 

Konkle, A. T., Keith, S. E., McNamee, J. P., & Michaud, D. (2017). Chronic noise exposure in the spontaneously hypertensive rat. Noise & health19(90), 213

 

Huo, R., Zeng, B., Zeng, L., Cheng, K., Li, B., Luo, Y., ... & Niu, R. (2017). Microbiota modulate anxiety-like behavior and endocrine abnormalities in hypothalamic-pituitary-adrenal axis. Frontiers in cellular and infection microbiology7, 489.

 

Li, Y. Q., Shrestha, Y., Pandey, M., Chen, M., Kablan, A., Gavrilova, O., ... & Weinstein, L. S. (2016). G q/11 α and G s α mediate distinct physiological responses to central melanocortins. The Journal of clinical investigation126(1), 40-49.

 

Ozone, C., Suga, H., Eiraku, M., Kadoshima, T., Yonemura, S., Takata, N., ... & Sasai, Y. (2016). Functional anterior pituitary generated in self-organizing culture of human embryonic stem cells. Nature communications7, 10351

 

Kondoh, K., Lu, Z., Ye, X., Olson, D. P., Lowell, B. B., & Buck, L. B. (2016). A specific area of olfactory cortex involved in stress hormone responses to predator odours. Nature532(7597), 103

 

Alghadir, A. H., & Gabr, S. A. (2015). Physical activity and environmental influences on adrenal fatigue of Saudi adults: biochemical analysis and questionnaire survey. Journal of physical therapy science27(7), 2045-2051.

 

Liang, Q., Zhong, L., Zhang, J., Wang, Y., Bornstein, S. R., Triggle, C. R., & Xu, A. (2014). FGF21 maintains glucose homeostasis by mediating the cross talk between liver and brain during prolonged fasting. Diabetes, 63(12), 4064-4075.

 

Sasaki, G., Zubair, M., Ishii, T., Mitsui, T., Hasegawa, T., & Auchus, R. J. (2014). The contribution of Serine 194 phosphorylation to steroidogenic acute regulatory protein function. Molecular Endocrinology, 28(7), 1088-1096.

 

Iijima, M., Yoshimizu, T., Shimazaki, T., Tokugawa, K., Fukumoto, K., Kurosu, S., & Chaki, S. (2014). Antidepressant and anxiolytic profiles of newly synthesized arginine vasopressin V1B receptor antagonists: TASP0233278 and TASP0390325. British journal of pharmacology, 171(14), 3511-3525. 

 

Jedel, S., Hoffman, A., Merriman, P., Swanson, B., Voigt, R., Rajan, K. B & Keshavarzian, A. (2014). A randomized controlled trial of mindfulness-based stress reduction to prevent flare-up in patients with inactive ulcerative colitis. Digestion, 89(2), 142-155.

 

Hueston, C. M., & Deak, T. (2014). The inflamed axis: the interaction between stress, hormones, and the expression of inflammatory-related genes within key structures comprising the hypothalamic–pituitary–adrenal axis. Physiology & behavior, 124, 77-91.

 

Blandino Jr, P., Hueston, C. M., Barnum, C. J., Bishop, C., & Deak, T. (2013). The impact of ventral noradrenergic bundle lesions on increased IL-1 in the PVN and hormonal responses to stress in male sprague dawley rats. Endocrinology, 154(7), 2489-2500.

 

Joseph, A., Tang, M., Mamiya, T., Chen, Q., Yang, L. L., Jiao, J., & Tang, Y. P. (2013). Temporal association of elevated cholecystokininergic tone and adolescent trauma is critical for posttraumatic stress disorder-like behavior in adult mice. Proceedings of the National Academy of Sciences, 110(16), 6589-6594.

 

Goggin, S. L., Labrecque, M. T., & Allan, A. M. (2012). Perinatal exposure to 50ppb sodium arsenate induces hypothalamic-pituitary-adrenal axis dysregulation in male C57BL/6 mice. Neurotoxicology, 33(5), 1338-1345.

 

Suga, H., Kadoshima, T., Minaguchi, M., Ohgushi, M., Soen, M., Nakano, T & Sasai, Y. (2011). Self-formation of functional adenohypophysis in three-dimensional culture. Nature, 480(7375), 57-62.

 

References/Citations:How the ACTH ELISA was used:
The hypothermic response to bacterial lipopolysaccharide critically depends on brain CB1, but not CB2 or TRPV1, receptors
Alexandre, A.S. et al., (2011) J Physiol. 589:2415
Measure the concentration of ACTH in plasma obtained from control rats (saline) and LPS-induced hyperthermia rats.
Thymocyte-Synthesized Glucocorticoids Play a Role in Thymocyte Homeostasis and Are Down-Regulated by Adrenocorticotropic Hormone
Shengjun Qiao et al., Endocrinology, Sep 2009; 150: 4163 - 4169.
Measure the concentration of ACTH in sera obtained from C57BL/6 wild-type mice and genetically modified mice.
Plasma Osteopontin Modulates Chronic Restraint Stress-Induced Thymus Atrophy by Regulating Stress Hormones: Inhibition by an Anti-Osteopontin Monoclonal Antibody
Kathryn X. Wang et al., J. Immunol., Feb 2009; 182: 2485 - 2491.
Measure the level of ACTH in plasma obtained from wild-type and knock-out BALB/c mice under chronic restraint stress conditions.
Complex Role of the Mitochondrial Targeting Signal in the Function of Steroidogenic Acute Regulatory Protein Revealed by Bacterial Artificial Chromosome Transgenesis in Vivo
Goro Sasaki et al., Mol. Endocrinol., Apr 2008; 22: 951 - 964.
Measure plasma ACTH levels of 8-wk old mice of different genotypes. Blood was collected by cardiac puncture and ACTH was measured in pg/ml units.

Inhibition of brain proinflammatory cytokine synthesis reduces hypothalamic excitation in rats with ischemia-induced heart failure
Yu-Ming Kang et al., Am J Physiol Heart Circ Physiol, Jul 2008; 295: H227 - H236.

Measure plasma ACTH levels in Sprague-Dawley rats. Rats were subject to vehicle treatment in heart failure vs SHAM models. Blood was collected in chilled EDTA tubes, separated and stored at -80C.
Electroacupuncture activates corticotrophin-releasing hormone-containing neurons in the paraventricular nucleus of the hypothalammus to alleviate edema in a rat model of inflammation
Aihui Li, et al. BMC Complement Altern Med. 2008; 8:20
Measure plasma ACTH levels in male Sprague-Dawley rats. Plasma was prepared from blood centrifuged at (1310 g) for 15 minutes at 4 C, collected and stored at -80 C. Rats underwent various experimental treatments including electroacupuncture and inflammation.
Lesions of the anteroventral third ventricle region exaggerate neuroendocrine and thermogenic but not behavioral responses to a novel environment
Douglas G. Whyte and Alan Kim Johnson Am J Physiol Regulatory Integrative Comp Physiol, Jan 2007; 292: R137 - R142.
Measure plasma ACTH levels of male Sprague Dawley rats. Blood was collected in EDTA collection tubes, centrifuged at 1,000 g for 15 minutes at 4C, collected and stored at -80C. Rats were studied in SHAM and anteroventral third ventricle (AV3V) ablation models.
Neuroendocrine profiling in inherited stress-induced arterial hypertension rat strain with stress-sensitive arterial hypertension
A L Markel et al., J. Endocrinol., Dec 2007; 195: 439 - 450.
Measure plasma ACTH levels in rats. Blood was collected in ice-cold EDTA-coated tubes, centrifuge and collected for storage at -70C. Rats investigated were ISIAH and WAG strains under stress conditions.
The Effects of SOM230 on Cell Proliferation and Adrenocorticotropin Secretion in Human Corticotroph Pituitary Adenomas
Dalia L. Batista et al., J. Clin. Endocrinol. Metab., Nov 2006; 91: 4482 - 4488.
Measure ACTH levels in media from the primary cell culture of human corticotroph adenomas obtained from Cushing's disease patients that underwent transsphenoidal surgery (tumor samples).

 

 

 

How It Works

How It Works: 

The ACTH Immunoassay is a two-site ELISA (Enzyme-Linked Immunosorbent Assay) for the measurement of the biologically active 39 amino acid chain of ACTH. A goat polyclonal antibody to human ACTH, purified by affinity chromatography, and a mouse monoclonal antibody to human ACTH are specific for well defined regions on the ACTH molecule. One antibody is prepared to bind only the C-terminal ACTH 34-39 and this antibody is biotinylated. The other antibody is prepared to bind only the mid-region and N-terminal ACTH 1-24 and this antibody is labeled with horseradish peroxidase (HRP) for detection.

ACTH ELISA

In this assay, calibrators, controls, or patient samples are simultaneously incubated with the enzyme labeled antibody and a biotin coupled antibody in a streptavidin-coated microplate well. At the end of the assay incubation, the microwell is washed to remove unbound components and the enzyme bound to the solid phase is incubated with the substrate, tetramethylbenzidine (TMB). An acidic stopping solution is then added to stop the reaction and converts the color to yellow. The intensity of the yellow color is directly proportional to the concentration of ACTH in the sample. A dose response curve of absorbance unit vs. concentration is generated using results obtained from the calibrators. Concentrations of ACTH present in the controls and patient samples are determined directly from this curve.