General species 1,25-Dihydroxyvitamin D3 (DHVD3) (Competitive EIA) ELISA Kit

Mfr.No.

CEA467Ge

Sensitivity

9.92pg/mL

Detection Range

24.69-2,000pg/mL

Reactivity

General

Assay Duration

Assay Type

Competitive Inhibition

Description

This assay has high sensitivity and excellent specificity for detection of 1,25-Dihydroxyvitamin D3 (DHVD3).
No significant cross-reactivity or interference between 1,25-Dihydroxyvitamin D3 (DHVD3) and analogues was observed.

Alternative Names

DH-VD3; Calcitriol; Rocaltrol; Calcijex; 1,25-(OH)2D3; 1,25-Dihydroxycholecalciferol; 1,25-Dihydroxy Vitamin D3

Detection Method

Colormetric

Kit Components

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

Recovery

Matrices listed below were spiked with certain level of 1,25-Dihydroxyvitamin D3 (DHVD3) and the recovery rates were calculated by comparing the measured value to the expected amount of 1,25-Dihydroxyvitamin D3 (DHVD3) in samples.

Matrix	Recovery range (%)	Average(%)
serum(n=5)	95-104	101
EDTA plasma(n=5)	83-95	87
heparin plasma(n=5)	86-98	95

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of 1,25-Dihydroxyvitamin D3 (DHVD3) 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)	93-101%	81-92%	88-98%	89-96%
EDTA plasma(n=5)	87-102%	92-101%	92-99%	92-101%
heparin plasma(n=5)	78-89%	93-101%	78-103%	96-103%

Features

Detects small molecule

Species

General species

Precautions

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.

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level 1,25-Dihydroxyvitamin D3 (DHVD3) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level 1,25-Dihydroxyvitamin D3 (DHVD3) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: C<10%
Inter-Assay: C<12%

Notes

This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to DHVD3 has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled DHVD3 analogues and unlabeled antigen (Standards or samples) with the pre-coated antibody. After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of DHVD3 in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of DHVD3 in the sample.

Usage Notes

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.

Protocol

Datasheet

Applications	ELISA
Research Area	Metabolic pathway

Storage	4°C for 1 month/ -20°C for long-term(within shelf life)
Shelf Life	one year

Reference	Selective Cytotoxic Action and DNA Damage by Calcitriol-Cu(II) Interaction: Putative Mechanism of Cancer Prevention Calcium-deficiency assessment and biomarker identification by an integrated urinary metabonomics analysis Calcitriol induced redox imbalance and DNA breakage in cells sharing a common metabolic feature of malignancies: Interaction with cellular copper (II) ions leads to the production of reactive oxygen species. Treatment of established left ventricular hypertrophy with fibroblast growth factor receptor blockade in an animal model of CKD Vitamin D Prevents Podocyte Injury via Regulation of Macrophage M1/M2 Phenotype in Diabetic Nephropathy Rats Impact of Curcumin and Flaxseed Oil on Bone Turnover in Glucocorticoid-Injected Female Rats Calcitriol–copper interaction leads to non enzymatic, reactive oxygen species mediated DNA breakage and modulation of cellular redox scavengers in hepatocellular …

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SKU	Supplier No.	Size	Your price
BHE10101782	CEA467Ge	96T*1	$796	Add to Cart
BHE10101782	CEA467Ge	96T*5	$3582	Add to Cart
BHE10101782	CEA467Ge	96T*10	$6766	Add to Cart

ELISA Kit for 1,25-Dihydroxyvitamin D3 (DHVD3) (General species)

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