- Overview
- Data/Specifications
- Literature/Support
- How It Works
- Related Products
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
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
Product Insert:
ACTH ELISA Insert (PDF)
Articles/FAQ:
Stress and Autoimmune Disease: the role of ACTH (blog post)
ELISA Data Reduction Guide
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da Silva Oliveira Barbosa, E., Roggero, E. A., González, F. B., Fernández, R. D. V., Carvalho, V. F., Bottasso, O. A., ... & Villar, S. R. (2020). Evidence in Favor of an Alternative Glucocorticoid Synthesis Pathway During Acute Experimental Chagas Disease. Frontiers in Endocrinology, 10, 866.
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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 & Development,33(3-4), 209-220.
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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 Endocrinology,470, 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 Foods,45, 181-189.
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| 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 JohnsonAm 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
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.
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.
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反应体系如下:
plasmid10ul
10xKbuffer5ul
KpnI1ul
BamHI1ul注意千万不可多加总酶量必须<4%
ddH20upto50ul
总体积改变加酶量按比例改变.
炎性体（inflammasome）是细胞内的一类多蛋白复合物，在炎性反应中发挥着至关重要的作用。炎性体包括半胱天冬酶-1（caspase1）、PYCARD和NALP，有时也包括半胱天冬酶-5（caspase5，也被称作半胱天冬酶-11或ICH-3）。它是在骨髓细胞（myeloidcell）中产生的，也是先天性免疫系统的一个组分。炎性体的确切组成依赖于启动炎性体组装的激活物，如双链RNA和石棉会引发不同的炎性体组成。炎性体促进炎性细胞因子IL-1β和IL-18成熟。
在一项新的研究中，来自比利时法兰德斯生物技术中心/根特大学（VIB/UGent）的LieselotteVandeWalle博士、DanielJiménezFernández以及教授MoLamkanfi研究团队对半胱天冬酶-12（caspase12）的功能产生新的认识。基于此，他们打破了这个领域对半胱天冬酶-12的固执观念：半胱天冬酶-12是炎性体的负调节物。这些新的认识为研究人员挣脱现有的研究路线和鉴定它的真正生理学功能铺平道路。相关研究结果发表在2016年6月2日那期Nature期刊上，论文标题为“Doescaspase-12suppressinflammasomeactivation?”。
研究人员也指出这将需要进行大量的“重新研究（re-researching）”。之前所谓的半胱天冬酶-12在细胞死亡、应激反应、疟疾和败血症等中的作用---引用了9000多次---必需复核，这是因为这些作用经常是基于不正确的小鼠模型得出的。
VIB/UGent教授MoLamkanfi说，“我们发现在很多情形下，对半胱天冬酶-12的研究是基于对半胱天冬酶-11（caspase11）和半胱天冬酶-12都进行基因敲除的小鼠模型开展的。因此从研究结果中推断半胱天冬酶-12的作用是不可能的。我们如今引入新的选择性半胱天冬酶-12基因敲除小鼠，这应当能够让我们追踪半胱天冬酶-12的确切功能。”
利用这些新的小鼠，Lamkanfi团队证实在体外模拟的BMDM（bonemarrow-derivedmacrophage,骨髓衍生性巨噬细胞）和体内接种的小鼠中，半胱天冬酶-12缺乏都不能增加半胱天冬酶-1激活。剔除半胱天冬酶-12也不会增强炎性体途径释放出成熟的IL-1β和IL-18。他们的发现表明不论半胱天冬酶-11的表达状态如何，半胱天冬酶-12都不会作为半胱天冬酶-1激活的生理学上负显性调节物，因而也不会作为炎性体的生理学上负显性调节物。
因为溶蛋白酶是蛋白质本质的酶
请教各位老师，
文献中检测细胞的活性有的用“NADPHdehydrogenase（NADPH脱氢酶）”有的用“NADPHdiaphorase（NADPH黄递酶）”
它们是同一种酶吗？它们的功能是什么？检测它们的活性是否能够评估细胞的活性。
不胜感激
