For quantitative determination of corticosterone in biological fluids such as plasma, serum or urine samples of mice, rats and other species, as well as in cells or tissue culture supernatant.

Corticosterone EIA


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Background

Corticosterone (C21H30O4, also called 11b,21-Dihydroxyprogesterone, Reichstein’s Substance H, or Kendall’s Compound B) is, like cortisol and cortisone, a glucocorticoid hormone secreted from the cortex of adrenal gland. Corticosterone is derived from cholesterol through a series of enzymatically mediated steps and also serving as a precursor of aldosterone. It is a primary glucocorticoid in mice and rats and other animals (such as rabbits, birds, amphibians, and reptilians) in which the 17a-hydroxylase is supposed not to exist in adrenal gland. Corticosterone is produced under the control of ACTH and the production has a circadian rhythm with peak levels in the latter portion of the day in nocturnal animals like rats and is believed to play a decisive role in sleep-wake cycles (1). Corticosterone can be used as a non-invasive biomarker of stress study through the collection of urine and feces to avoid corticosterone increase of blood levels which is caused by normal invasive methods (2). Corticosterone is also being studied in different fields such as impairment of long-term memory retrieval (3), chronic corticosterone elevation due to dietary restrictions (4) and response to burn injuries (5) etc.

Since most of corticosterone in blood is bound to a plasma protein called corticosterone- binding globulin (CBG), the determination of blood corticosterone with presently available commercial assay kits requires an initial extraction procedure. On the other hand, the present assay kit for corticosterone newly developed by our laboratory provides a tool for direct determination of corticosterone in blood by simple dilution of blood samples with the diluent included in the kit. Furthermore, assays using the kit can be completed within a short period. The corticosterone EIA kit newly developed will be a quite useful tool for further development of corticosterone research.

 

Biomarkers@flyer
Biomarker mesurement kit Flyer download [PDF]

Feature and Advantages

This EIA kit is used for quantitative determination of corticosterone in biological fluids such as plasma, serum or urine samples of mice, rats and other species, and also cell or tissue culture supernatant. It has various advantages, such as no extraction procedure of samples, short assay time, practically no influences of other body fluids or physiological active substances coexisting in samples assayed.

EThe assay kit range: 0.21-50ng/mL.
EThe assay running time: 2h. + 0.5 h.
EMaximum measurable samples: 41 in duplicate
ETest sample: plasma, serum, urine, and cell or tissue culture supernatant etc.
EThe 96-well plate in the kit is consisted of 8-wells strips, so that the kit can be used dividedly in strips.
EPrecision and reproducibility

  Intra-assay %CV: 2.5~4.7

  Inter-assay %CV: 7.5~9.8.

EStability and Storage Store all the components in the kit at 2-8Ž

  The expiry date is stated on the package.

 

<Specificity>
 Cross reactivities of the antibody used in the kit.

CompoundCross Reactivity i“j 
Corticosterone 100
11-Deoxycorticosterone 15.5
Progesterone <5.9
Androstenedione <5.4
Testosterone <3.9
Aldosterone <1.7
Cortisol <0.5
Cortisone <0.4
Prednisolone <0.34
Betamethasone <0.1
Dexamethasone <0.09
Triamcinolone <0.08 
Predmisone  <0.06 
Pregnenolone  <0.06 
DHEA <0.05
Estradiol 0
Cholesterol 0

 

<Principle>
This EIA kit for determination of corticosterone is based on a competitive enzyme immunoassay using combination of specific antibody to corticosterone and corticosterone- horseradish peroxidase (HRP) conjugate (HRP-labeled corticosterone) system. The 96 wells plate is coated with goat anti rabbit IgG, to which corticosterone standard or samples, HRP-labeled corticosterone and specific antibody are added for competitive immunoreaction. After incubation and plate washing, HRP enzyme activity is determined by 3,3’,5,5’- tetramethylbenzidine (TMB) and the concentration of corticosterone is calculated.

Kit component

 ComponentFormQuantityMain Ingredient
1 Antibody Coated Plate microtiter plate 1 plate (96 wells) Goat anti rabbit IgG
2 Corticosterone Standard lyophilized powder 1 vial (50ng) Synthetic corticosterone
3 HRP-Labeled Corticosterone liquid 1 vial (0.3mL) HRP conjugated corticosterone
4 Specific Antibody liquid 1 bottle (7 mL) Rabbit anti corticosterone antibody
5 TMB Substrate liquid 1 bottle (12 mL) 3,3’,5,5’-Tetramethylbenzidine (TMB)
6 Reaction Stopping Solution liquid 1 bottle (12 mL) 1M Sulfuric acid
7 Buffer Solution liquid 1 bottle (10 mL) BSA-containing PBS buffer
8 Sample Diluent liquid 1 bottle (50 mL) A specially formulated displacer of CBG
9 Concentrated Wash Solution liquid 1 bottle (25 mL) Concentrated saline
10 Adhesive Foil   2 sheets  
References

1. Vázquez-Palacios G, Retana-Márquez S et al: Further definition of the effect ofcorticosterone on the sleep-wake pattern in the male rat. Pharmacol Biochem Behav 70:305-310, 2001
2. Goymann W, Möstl E and Gwinner E: Corticosterone metabolites can be measured noninvasively in excreta of European Stonechats (Saxicola torquata rubicola). The Auk119:1167-1173. 2002
3. Hupe JM, James AC et al: Cortical feedback improves discrimination between figure and background by V1, V2 and V3 neurons. Nature 394:784-787, 1998
4. Kitaysky AS, Kitaiskaiya EV et al: Dietary restriction causes chronic elevation of corticosterone and enhances stress response in red-legged kittiwake chicks.  J Comp Physiol B:Regulatory, Integrative and Comparative Physiology 171:701-709, 2001
5. Thelling O, Noel G et al: Stress hormone secretion and gut signal transducer (STAT) proteins after burn injury in rats. Shock 16:393-397, 2001
6. National Committee for Clinical Laboratory Standards Evaluation Protocols, SC1 (1989), Vallanova, PA: NCCLS
7. Takaya J, Iharada A, et al: Upregulation of Hepatic 11_-Hydroxysteroid Dehydrogenase-1
Expression in Calcium-Deficient Rats. Ann Nutr Metab. 59:73-78, 2011
8. Satoh S, Suzuki H et al: Casted-immobilization downregulates glucocorticoid receptor expression in rat slow-twitch soleus muscle. Life Sciences. 89:962–967, 2011
9. Ekuni D, Tomofuji K et al: Occlusal disharmony increases amyloid-_ in the rat hippocampus,
NeuroMolecular Medicine, 13:197-203, 2011
10. Irie K, Ekuni D et al: Occlusal disharmony induces BDNF level in rat submandibular gland,
Archives of Oral Biology. 56:35-40, 2011
11. Yamawaki Y, Fuchikami M et al: Antidepressant-like effect of sodium butyrate (HDAC inhibitor) and its molecular mechanism of action in the rat hippocampus. World Journal of Biological Psychiatry. 2011 Aug 3. [Epub ahead of print]
12. Takei Y, Bartolo RC et al: Water deprivation induces appetite and alters metabolic strategy in Notomys alexis: unique mechanisms for water production in the desert. Proc Bio Sci, 2012 Mar 7. [Epub ahead of print]
(Remark: The references of no. 7-12 are publications used of this product in their studies.)

Product List

Species: mice, rats and other species 
Assay range: 0.21-50ng/mL
Assay sample volume: 100 µL (Plasma, Serum, urine, culture supernatant)
Assay time: 2h. +0.5 hr.


Product Name Cat# Quantity Price

Corticosterone EIA DataSheet ,

YII-YK240-EX

1 KIT ¥76,000
$1014

Product Name Cat# Quantity Price

Corticosterone (Compd. B)-3-CMO-HRP, Horseradish Peroxidase DataSheet

FKA-419

1000 TEST ¥25,000
$334

Anti Corticosterone-3-CMO, - (Rabbit)  DataSheet

FKA-420-E

2000 TEST ¥25,000
$334

Anti Corticosterone, - (Rabbit)  DataSheet

FKA-420

2000 TEST ¥25,000
$334

Deoxycorticosterone (DOC)-3-HRP, Horseradish Peroxidase

FKA-421

1000 TEST ¥25,000
$334

Anti DOC, - (Rabbit) Unlabeled

FKA-422-E

2000 TEST ¥25,000
$334

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