Intended Use
Enzyme immunoassay for the quantitative measurement of total 25-OH-Vitamin D (Vitamin D2 and D3) in human serum. For research use only. Not for use in diagnostic procedures.
Storage
The kit is shipped at ambient temperature and should be stored at 2-8 °C. Keep away from heat or direct sunlight. The storage and stability of specimens and prepared reagents is stated in the corresponding chapters.
The microtiter strips are stable up to the indicated expiry after the kit is opened. Make sure that the opened bag is tightly closed when stored at 2-8 °C.
Precision
Performance parameters were measured according to Clinical & Laboratory Standards Institute (CLSI) guidelines.
The Intra-Assay and Inter-Assay study was conducted during 20 days using one reagent lot. Two runs were performed per day with high and low kit controls and with a panel of five human serum samples. Each sample was run in duplicate. The results were calculated according CLSI-EP05-A3.
The intra-assay precision showed a mean CV from 5.9% and a range of 3.2% - 11.9% for the five different human serum samples.
The inter-assay precision showed a mean CV from 8.3% and a range of 5.3% - 15.6% for the five different human serum samples.
The between lot variation study was conducted during 5 days of testing. Each run were performed per day with high and low kit controls and with a panel of five human serum samples. Each sample was tested in five replicate per run with 3 different reagent lots. The results were calculated according CLSI-EP05-A3.
The between lot variation showed a mean CV from 10.7% and a range of 6.1%– 20.1% for the five different human serum samples.
Detection Limit
Limit of Blank (LoB)
The LoB study was conducted during three days of testing by two operators. The runs were performed with high and low kit controls and with five different blank samples. For each sample four replicates were tested using two reagent lots.
Limit of Blank = 6.2 ng/mL
Limit of Detection (LoD)
The LoD study was conducted during three days of testing by two operators. The runs were performed with high and low kit controls and with five different low concentrated human serum samples. For each sample four replicates were tested using two reagent lots.
Limit of Detection = 11.6 ng/mL
Limit of Quantitation (LoQ)
The LoQ study was conducted during three days of testing by two operators. The runs were performed with high and low kit controls and seven different low to mid concentrated human serum samples. For each sample four replicates were tested using two reagent lots.
Limit of Quantitation = 8.2 ng/mL
General Description
Vitamin D refers to a group of fat-soluble secosteroids being responsible for intestinal absorption of calcium, iron, magnesium, phosphate and zinc. The most important vitamin Ds in humans are vitamin D3 (cholecalciferol) and vitamin D2 (ergocalciferol). Vitamin D3 is synthesized in the skin from the cholesterol precursor 7-dehydrocholesterol and is the major source of vitamin D in humans. In the liver vitamin D is further metabolized via hydroxylation to 25-hydroxyvitamin D (25-OH-vitamin D). 25-OH-vitamin D is the major circulating metabolite of vitamin D. 25-OH-vitamin D is the precursor for other vitamin D metabolites, but shows also limited activity by itself.
Citations
Publication ()
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Vitamin D mitigates diabetes-associated metabolic and cognitive dysfunction by modulating gut microbiota and colonic cannabinoid receptor 1
Hussein HM, Elyamany MF, Rashed LA, Sallam NA.
Applications: ELISA
Reactive species: Rat
"Abstract: Introduction: Obesity is associated with elevated endocannabinoid tone, gut dysbiosis, and inflammation predisposing to diabetes. The endocannabinoid system mediates the effects of gut microbiota and regulates the gut barrier integrity. We examined the effects of vitamin D (VD) on colonic cannabinoid receptor 1(CB1R), tight junction proteins, gut dysbiosis, metabolic and cognitive dysfunction in a model of type 2 diabetes compared with metformin."
Article snippet: Serum VD level was determined by an ELISA kit, catalog # DEIA2219 (Creative Diagnostics, USA). 
Figure 1. Effects of VD and metformin on serum VD levels in HFSD-fed/STZ-treated rats.
Active form of vitamin D analogue mitigates neurodegenerative changes in Alzheimer's disease in rats by targeting Keap1/Nrf2 and MAPK-38p/ERK signaling pathways
Saad El-Din S, Rashed L, Medhat E, Emad Aboulhoda B, Desoky Badawy A, Mohammed ShamsEldeen A, Abdelgwad M
Applications: ELISA
Reactive species: Rat
"Abstract: Background: The nuclear factor erythroid2-related factor2 (Nrf2), a chief transcriptional regulator of antioxidant response element (ARE), is considered a promising target for the prevention of Alzheimer's disease (AD). Vitamin D has been recognized to have a crucial role in improving AD cognitive functions. The present study was conducted to evaluate the effects of active vitamin D analogue, Maxacalcitol, on Keap1-Nrf2 signaling pathway in experimental Alzheimer's disease in rats.Materials and methods:The study was conducted on thirty female white albino rats divided equally into 3 groups: control group, Alzheimer group induced by Lipopolysaccharide and Alzheimer group treated with active vitamin D3 analogue, Maxacalcitol. The following parameters were assessed in rat brain tissues: Gene expression of Nrf2, Keap1 and MAF by RT-PCR, protein levels of phosphorylated MAPK-38p and ERK1/2 by Western Blot Technique, estimation of HO-1, Amyloid β, p-Tau levels and serum levels of TNFα, IL-10 and total 25-hydroxyvitamin D, serum calcium levels, GSH and MDA levels were also estimated in addition to cognitive function tests and histo-pathological examination of rat brain tissues."
Article snippet: Estimation of serum levels of total 25-hydroxyvitamin D by ELISA (Cat.No: DEIA2219, Creative Diagnostics, NY, USA) according to the manufacturer‘s recommendations.
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