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You are here:Home » Kits and Assays » Kits for ELISA (Hormones & Steroids), BioAssay™ » Adrenocorticotropic Hormone (ACTH) BioAssay™ ELISA Kit (Rat)

Adrenocorticotropic Hormone (ACTH) BioAssay™ ELISA Kit (Rat)

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Specifications

Adrenocorticotropic Hormone (ACTH) BioAssay™ ELISA Kit (Rat) is a competitive inhibition enzyme immunoassay technique for the in vitro quantitative measurement of ACTH in rat serum, plasma and other biological fluids.
Catalog #023173
Adrenocorticotropic hormone (ACTH), also known as corticotropin, is a polypeptide tropic hormone produced and secreted by the anterior pituitary gland. It is an important component of the hypothalamic-pituitary-adrenal axis and is often produced in response to biological stress (along with its precursor corticotropin-releasing hormone from the hypothalamus). Its principal effects are increased production and release of corticosteroids. A deficiency of ACTH is a cause of secondary adrenal insufficiency and an excess of it is a cause of Cushing's disease.
Pro-opiomelanocortin (POMC) is a precursor polypeptide with 241aa residues. It is synthesized by corticotrope cells of the anterior pituitary gland, melanotrope cells of the intermediate lobe of the pituitary gland, about 3000 neurons in the arcuate nucleus of the hypothalamus, and smaller populations of neurons in the dorsomedial hypothalamus and brainstem. The large molecule of POMC is the source of several important biologically active substances. POMC can be cleaved enzymatically into the following peptides adrenocorticotrophic hormone (ACTH) and gamma LPH in the anterior pituitary gland CLIP, gamma LPH, gamma MSH and beta-endorphin in the intermediate lobe gamma-MSH met-enkephalin.
Each of these peptides is packaged in large dense-core vesicles that are released from the cells by exocytosis in response to appropriate stimulation. gamma MSH produced by neurons in the arcuate nucleus has important roles in the regulation of appetite and sexual behavior, while alpha-MSH secreted from the intermediate lobe of the pituitary regulates the production of melanin. ACTH is a peptide hormone that regulates the secretion of glucocorticoids from the adrenal cortex. beta-endorphin and met-enkephalin are endogenous opioid peptides with widespread actions in the brain.
Detection Range12.35-1000pg/ml
Sensitivity4.39pg/ml
Intra-AssayCV 10%
Inter-AssayCV: 12%
Kit Components*023173A: Microtiter Plate, 1x96wells, Pre-coated, ready to use
*023173B: Standard, 2x1vial
023173C: Standard Diluent, 1x20ml
*023173D: Detection Reagent A (green), 1x120ul
*023173E: Detection Reagent B (red), 1x120ul
023173F: Assay Diluent A, 1x12ml
023173G: Assay Diluent B, 1x12ml
023173H: TMB Substrate, 1x9ml
023173K: Stop Solution, 1x6ml
023173L: Wash Buffer, 30X, 1x20ml
PrecautionThe Stop Solution (023173K) suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.
Storage and StabilityStore *023173A, *023173B, *023173D and *023173E at -20°C. Store all the other components at 4°C. Unused kit is stable for 6 months after receipt. Once kit components are opened, it is highly recommended to use remaining reagents within 1 month provided this is within the expiration date of the kit. For maximum recovery of product, centrifuge the original vials after thawing and prior to removing the cap.
Protocol Summary1. Prepare all reagents, samples and standards.
2. Add 50ul standard, control or sample to each well.
3. Add 50ul prepared Detection Reagent A immediately. Shake and mix. Incubate 1 hour at 37°C.
4. Aspirate and wash 3 times.
5. Add 100ul prepared Detection Reagent B. Incubate 30 minutes at 37°C.
6. Aspirate and wash 5 times.
7. Add 90ul 023173H: TMB Substrate. Incubate 15-25 minutes at 37°C.
8. Add 50ul 023173K: Stop Solution. Read at 450nm immediately.
Important NoteThis product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.


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