Allopregnanolone EIA Kit (One Plate)

Allopregnanolone EIA Kit (One Plate)

Article No K044-H1
Product Type Assay & Detection
Application ELISA
Size 1x96 well plate

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Specifications

Country Availability DK, EE, FI, IS, LT, LV, NO, SE
Data Sheet Link https://www.arborassays.com/documentation/inserts/K044-H.pdf
Description Allopregnanolone EIA Kit (One Plate)
Supplier Arbor Assays Inc.
Notes Allopregnanolone (3 a -hydroxy-5 a -pregnan-20-one) is a neurosteroid present in the blood and also the brain. Allopregnanolone is made from progesterone which is converted into 5 a -dihydroprogesterone by 5 a -reductase type I. 3 a - hydroxysteroid oxidoreductase isoenzymes converts this intermediate into allopregnanolone. 3cr-hydroxysteroids do not interact with classical intracellular steroid receptors but bind stereoselectively and with high affinity to receptors for the major inhibitory neurotransmitter in brain, g -amino-butyric acid (GABA) (1). While allopregnanolone, like other GABA A receptor active neurosteroids, such as allotetrahydrodeoxycorticosterone, positively modulates all GABA A receptor isoforms, those isoforms containing δ -subunits exhibit greater magnitude potentiation. As physiologic consequences, it may be involved in neuronal plasticity, learning and memory processes, aggression and epilepsy, and the modulation of the responses to stress, anxiety and depression. Allopregnanolone has pharmacological properties similar to other positive modulators of GABA A receptors, including anxiolytic and anticonvulsant activity (2). Anxiety and depression are common side effects of 5 a -reductase inhibitors such as finasteride and dutasteride, and they are believed to be caused, in part, by the prevention of the endogenous production of allopregnanolone.
Protocol The DetectX® Allopregnanolone Immunoassay kit is designed to quantitatively measure Allopreg - nanolone present in serum, plasma, extracted dried fecal samples, urine and tissue culture media samples. Please read the complete kit insert before performing this assay. An allopregnanolone standard is provided to generate a standard curve for the assay and all samples should be read off the standard curve. Standards or diluted samples are pipetted into a clear microtiter plate coated with an antibody to capture rabbit antibodies. An allopregnanolone-peroxidase conjugate is added to the standards and samples in the wells. The binding reaction is initiated by the ad - dition of a polyclonal antibody to allopregnanolone to each well. After a 2 hour incubation the plate is washed and substrate is added. The substrate reacts with the bound allopregnanolone- peroxidase conjugate. After a short incubation, the reaction is stopped and the intensity of the generated color is detected in a microtiter plate reader capable of measuring 450nm wavelength. The concentration of the allopregnanolone in the sample is calculated, after making suitable cor - rection for the dilution of the sample, using software available with most plate readers.
References Allopregnanolone aids neurogenesis and has been found to reverse neuron proliferative deficit and cognitive deficits in mouse model of Alzheimer’s disease (3). Allopregnanolone has also been shown to restore functionality in a mouse model of Parkinson’s Disease (4). It has also been shown to improve behavioral problems in post-traumatic stress disorder (5,6). 1. Paul SM, Purdy RH., “Neuroactive Steroids”, FASEB J., 1992, 6:2311-2322. 2. Kokate TG, Svensson BE, and Rogawski MA. “Anticonvulsant activity of neurosteroids: correlation with gamma-aminobutyric acid-evoked chloride current potentiation.” J. Pharmacol. Exp. Ther. 1994, 270:1223-1229. 3. Wang JM., et al ., “Allopregnanolone reverses neurogenic and cognitive deficits in mouse model of Alzheimer’s disease.” PNAS. 2010, 107:6498–6503. 4. Adeosun, SO, et. al, “Allopregnanolone Reinstates Tyrosine Hydroxylase Immunoreactive Neurons and Motor Performance in an MPTP-Lesioned Mouse Model of Parkinson’s Disease”, 2012, PLOS One, 7(11):e50040. 5. Brunton, PJ, et. al,, “Central Opioid Inhibition of Neuroendocrine Stress Responses in Pregnancy in the Rat Is Induced by the Neurosteroid Allopregnanolone”. 2009, j. Neurosci., 29:6449-6460. 6. Pinna, G, “In a mouse model relevant for PTSD, selective brain steroidogenic stimulants (SBSSs) improve behavioral deficits by normalizing allopregnanolone biosynthesis” Behav Pharmacol. 2010, 21: 438-450.
Sensitivity 0.13 ng/mL
Species Reactivity species independent
Storage This kit should be stored at -20°C until the expiration date of the kit. The kit can be stored at 4°C up to the expiration date on the kit label, except for the Allopregnanolone Conjugate which must be stored at -20°C. The Allopregnanolone Conjugate can be freeze-thawed multiple times.
Technical Specifications Contents:Coated Clear 96 Well Plates Clear plastic microtiter plate(s) coated with goat anti-rabbit IgG. Kit K044-H1 Or -H5 1 Or 5 Each Catalog Number X016-1EA Allopregnanolone Standard Allopregnanolone at 1,000 ng/mL in a special stabilizing solution. Kit K044-H1 Or -H5 125 Or 625 μL Catalog Number C155-125UL Or -625UL DetectX ® Allopregnanolone Antibody A rabbit polyclonal antibody specific for allopregnanolone. Kit K044-H1 Or -H5 3 mL Or 13 mL Catalog Number C153-3ML Or -13ML DetectX ® Allopregnanolone Conjugate Allopregnanolone-peroxidase conjugate in a special stabilizing solution. Kit K044-H1 Or -H5 3 mL Or 13 mL Catalog Number C154-3ML Or -13ML Assay Buffer (or Concentrate) One plate kit uses a ready-to-use Assay Buffer. Five plate kit uses a 5X concentrate that should be diluted with deionized or distilled water. Kit K044-H1 28 mL Catalog Number X066-28ML Kit K044-H5 55 mL (Conc) Catalog Number X067-55ML Dissociation Reagent Kit K044-H1 Or -H5 1 mL Or 5 mL Catalog Number X058-1ML Or -5ML Dissociation Reagent is to be used only with Serum and Plasma samples. Wash Buffer Concentrate A 20X concentrate that should be diluted with deionized or distilled water. Kit K044-H1 Or -H5 30 mL Or 125 mL Catalog Number X007-30ML Or -125ML TMB Substrate Kit K044-H1 Or -H5 11 mL Or 55 mL Catalog Number X019-11ML Or -55ML Stop Solution A 1M solution of hydrochloric acid. CAUSTIC. Kit K044-H1 Or -H5 5 mL Or 25 mL Catalog Number X020-5ML Or -25ML Plate Sealer Kit K044-H1 Or -H5 1 Or 5 Each Catalog Number X002-1EA
Product Page Updated 2019-04-15T10:02:09.380Z

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Species reactivity:

species independent