ELISA Kit for Adrenocorticotropic Hormone (ACTH)

Corticotropin; Adrenocorticotropin

Specificity

This assay has high sensitivity and excellent specificity for detection of Adrenocorticotropic Hormone (ACTH).
No significant cross-reactivity or interference between Adrenocorticotropic Hormone (ACTH) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Adrenocorticotropic Hormone (ACTH) and the recovery rates were calculated by comparing the measured value to the expected amount of Adrenocorticotropic Hormone (ACTH) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 78-105 91
EDTA plasma(n=5) 88-97 93
heparin plasma(n=5) 97-105 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Adrenocorticotropic Hormone (ACTH) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Adrenocorticotropic Hormone (ACTH) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Adrenocorticotropic Hormone (ACTH) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 87-101% 81-103% 79-89% 78-97%
EDTA plasma(n=5) 80-96% 90-99% 80-101% 90-103%
heparin plasma(n=5) 78-97% 86-98% 78-103% 80-91%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

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Magazine Citations
Respir Res. Intracerebroventricular injection of leukotriene B4 attenuates antigen-induced asthmatic response via BLT1 receptor stimulating HPA-axis in sensitized rats. PubMed: 20403205
Toxicology Letters Regulation of corticosterone secretion is modified by PFOS exposure at different levels of the hypothalamic–pituitary–adrenal axis in adult male rats Pubmed:24440345
Mol Med Rep. Ursolic acid suppresses growth and adrenocorticotrophic hormone secretion in AtT20 cells as a potential agent targeting adrenocorticotrophic hormone-producing pituitary adenoma. Pubmed:24682498
Peptides Differential stress response in rats subjected to chronic mild stress is accompanied by changes in CRH-family gene expression at the pituitary level Pubmed:25236411
Journal of Animal and Feed Sciences Interdependence of the peripheral metabolism of glucocorticoids and thyroid hormones under calorie deficit in rats at different ages Researchgate:Source
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Cell Tissue Res CXCL10/CXCR3 signaling mediates inhibitory action by interferon-gamma on CRF-stimulated adrenocorticotropic hormone (ACTH) release PubMed: 26572542
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Sci Rep. Hedgehog signaling activation induces stem cell proliferation and hormone release in the adult pituitary gland Pubmed:27109116
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brain research bulletin Activation of ATP-sensitive potassium channel by iptakalim normalizes stress-induced HPA axis disorder and depressive behaviour by alleviating inflammation and oxidative stress in mouse hypothalamus. pubmed:28161195
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Toxicology Mixture effects of azole fungicides on the adrenal gland in a broad dose range pubmed:28454766
Biomedicine & Pharmacotherapy Triptolide suppresses growth and hormone secretion in murine pituitary corticotroph tumor cells via NF-kappaB signaling pathway pubmed:28892788
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Molecular and Cellular Endocrinology Metformin suppresses growth and adrenocorticotrophic hormone secretion in mouse pituitary corticotroph tumor AtT20 cells 10.1016:j.mce.2018.07.007
Frontiers in Behavioral Neuroscience Chronic hippocampal abnormalities and blunted HPA axis in an animal model of repeated unpredictable stress Pubmed:30079015
Placenta Human placentophagy: Effects of dehydration and steaming on hormones, metals and bacteria in placental tissue Pubmed:29941176
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