Antigen InformationAccession Number:P01193Gene ID:5443Gene Symbols:POMCProtein Name & Synonyms:Pro-opiomelanocortin (POMC) (Corticotropin-lipotropin) [Cleaved into: NPP; Melanotropin gamma (Gamma-MSH); Potential peptide; Corticotropin (Adrenocorticotropic hormone) (ACTH); Melanotropin alpha (Alpha-MSH); Corticotropin-like intermediary peptide (CLIP); Lipotropin beta (Beta-LPH); Lipotropin gamma (Gamma-LPH); Melanotropin beta (Beta-MSH); Beta-endorphin; Met-enkephalin]Assay FormatDetection Range:0.1-1,000 ng/ml Recommended Dilution (serum & plasma):Human: 2X / Mouse: 2X / Rat: 2XSensitivity:0.5 ng/mlSpecificity:This EIA kit is designed to detect human, mouse, and rat ACTHNumber of Targets Detected:1Species Detected:HumanMouseRatCompatible Sample Types:Cell Culture SupernatantsSerumDesign Principle:Competition-basedMethod of Detection:ColorimetricQuantitative/Semi-Quantitative:QuantitativeSolid Support:96-well MicroplateProduct SpecificationsSize:1, 2, or 5 x 96-Well Strip Microplate KitIntroductionAdrenocorticotropic hormone (ACTH, also adrenocorticotropin, corticotropin) is a 39 amino acids polypeptide tropic hormone. ACTH is synthesized from pre-pro-opiomelanocortin (pre-POMC), which is proteolytically cleaved by endopeptidases to yield various polypeptide fragments with varying physiological activity. ACTH is secreted by the anterior pituitary gland. It stimulates secretion of glucocorticoid steroid hormones from adrenal cortex cells, especially in the zona fasciculata of the adrenal glands. It influences steroid hormone secretion by both rapid short-term mechanisms and long-term actions. ACTH is also used as a medication and diagnostic agent.Product FeaturesStrip plates and additional reagents allow for use in multiple experimentsQuantitative protein detectionEstablishes normal rangeThe best products for confirmation of antibody array dataApplication NotesKit ComponentsPre-Coated 96-well Strip MicroplateWash BufferStandard PeptideAssay Diluent(s)Biotinylated PeptideHRP-StreptavidinTMB One-Step SubstrateStop SolutionAssay DiagramPositive Control SampleCapture AntibodyUser ManualOther Materials RequiredDistilled or deionized waterPrecision pipettes to deliver 2 µl to 1 ml volumesAdjustable 1-25 ml pipettes for reagent preparation100 ml and 1 liter graduated cylindersTubes to prepare standard and sample dilutionsOrbital shakerAluminum foilSaran WrapAbsorbent paperMicroplate reader capable of measuring absorbance at 450nmSigmaPlot software (or other software that can perform four-parameter logistic regression models)Protocol OutlinePrepare all reagents, samples and standards as instructed.Add 100 µl detection antibody to each well.Incubate 1.5 h at RT or O/N at 4°C.Add 100 µl standard or sample to each well.Incubate 2.5 h at RT.Add 100 µl prepared streptavidin solution.Incubate 45 min at RT.Add 100 µl TMB One-Step Substrate Reagent to each well.Incubate 30 min at RT.Add 50 µl Stop Solution to each well.Read plate at 450 nm immediately.Storage/StabilityStandard, Biotinylated ACTH peptide, and Positive Control should be stored at -20°C after arrival. Avoid multiple freeze-thaws. The remaining kit components may be stored at 4°C. Opened Microplate Wells and antibody (Item N) may be stored for up to 1 month at 2° to 8°C. Return unused wells to the pouch containing desiccant pack and reseal along entire edge.We used a serum dilution of 1:4. Works great.AnonymousKent State01/03/2020, 03:31 PMThis works well. I did 5 fold dilution with mouse serum and still worked.AnonymousYale University10/29/2019, 02:21 PMThis kit works well and produces consistent results for mouse serum. We used a serum dilution of 1:4 or 1:8 with a total reaction volume of 100uL.AnonymousThe Ohio State University03/20/2019, 09:13 PMWrite A ReviewManualSDS