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25(OH) Vitamin D ELISA kit is a Competitive ELISA kit for the measurement of 25(OH) Vitamin D in Human in Plasma, Serum samples.

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Key facts

Detection method

Colorimetric

Sample types

Plasma, Serum

Assay type

Competitive

Reactive species

Human

Range

0.5 - 1010 ng/mL

Assay time

1h 30m

Sensitivity

= 1.98 ng/mL

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Reactivity data

Application

cELISA

Reactivity

Reacts

Dilution info

-

Notes

-

What's included?

1 x 96 Tests
Components
25(OH) Vitamin D Antibody
1 x 5 mL
25(OH) Vitamin D Conjugate (100X)
1 x 50 µL
25(OH) Vitamin D Standard 1 (1010 ng/mL)
1 x 40 µL
25(OH) Vitamin D Standard 2 (279 ng/mL)
1 x 40 µL
25(OH) Vitamin D Standard 3 (71.6 ng/mL)
1 x 40 µL
25(OH) Vitamin D Standard 4 (24.4 ng/mL)
1 x 40 µL
25(OH) Vitamin D Standard 5 (4.8 ng/mL)
1 x 40 µL
25(OH) Vitamin D Standard 6 (0.5 ng/mL)
1 x 40 µL
25(OH) Vitamin D3 Conjugate Diluent
1 x 6 mL
Dissociation Buffer
1 x 10 mL
Donkey anti-sheep IgG Microplate (12 x 8 wells)
1 x 96 Test
Plate Sealer
3 x 1 Unit
Sample Diluent
1 x 1.8 mL
Stop Solution
1 x 5 mL
Wash Buffer 4 Concentrate (20X)
1 x 20 mL
pNpp Substrate
1 x 20 mL

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25(OH) Vitamin D ELISA kit is a Competitive ELISA kit for the measurement of 25(OH) Vitamin D in Human in Plasma, Serum samples.

Key facts

Detection method

Colorimetric

Sample types

Plasma, Serum

Assay type

Competitive

Reactive species

Human

Range

0.5 - 1010 ng/mL

Assay time

1h 30m

Assay Platform

Pre-coated microplate (12 x 8 well strips)

Sensitivity

= 1.98 ng/mL

Precision

Intra assay

Sample

Sample 1

n

20

mean

225.8 ng/mL

SD

-

C.V.

1.6

Sample

Sample 2

n

20

mean

37.3 ng/mL

SD

-

C.V.

2.1

Sample

Sample 3

n

20

mean

5.3 ng/mL

SD

-

C.V.

3.4

Inter assay

Sample

Sample 1

n

30

mean

297.2 ng/mL

SD

-

C.V.

11.5

Sample

Sample 2

n

30

mean

38.4 ng/mL

SD

-

C.V.

15.8

Recovery

Sample specific recovery

Sample type

Serum

Average %

= 91.9

Range

84 - 97.9 %

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

-20°C

Appropriate long-term storage conditions

-20°C

Storage information

-20°C

Notes

25(OH) Vitamin D ELISA kit is a complete kit for the quantitative determination of 25(OH) Vitamin D3 and 25(OH) Vitamin D2 and was tested in human plasma and serum samples.

It is recommended that you read the entire kit insert before proceeding with the assay.

Recent research efforts have shown that Vitamin D levels affect various disease states and are being linked with numerous indicators of well-being in humans. These include bone diseases such as osteoporosis and arthritis, but also additional disease including hypertension, diabetes, cancer and heart disease to name a few. Our Vitamin D ELISA kit offers an alternative to labor intensive and/or costly methods of testing for Vitamin D levels in human plasma and serum.

The transformation to the active form of Vitamin D begins with 7-dehydrocholesterol being acted upon by UV rays from the sun to form parent Vitamin D3. Alternatively, Vitamin D can be ingested as parent Vitamin D2 from various food sources, native or fortified. These parent compounds are transported to the liver and undergo hydroxylation to 25(OH) Vitamin D. This metabolite is then transported to the kidney where it undergoes a second hydroxylation to 1,25(OH)2 Vitamin D, the biologically active form of Vitamin D. It is important to note that levels of Vitamin D metabolites increase proportionately with increased uptake of parent Vitamin D. This combined with the greater half-life and stability of 25(OH) Vitamin D in circulation versus the active form (25 days versus 8 hours) are the reasons that the detection of the 25(OH) Vitamin D metabolite is used as the indicator for total Vitamin D concentration.

We have received feedback that this kit is not compatible with mouse samples.

Dissociation Buffer is added to wells coated with donkey anti-sheep IgG antibody. Standards and Samples are then added to these wells and dissociation of 25(OH) Vitamin D from Vitamin D Binding protein allowed to occur.

A solution of alkaline phosphatase conjugated 25(OH) Vitamin D3 is then added, followed by a solution of sheep monoclonal antibody to 25(OH) Vitamin D.

During incubation at room temperature the antibody binds the 25(OH) Vitamin D from the sample/standard or conjugate in a competitive manner and is itself captured by the anti-sheep IgG antibody. The plate is then washed, leaving a complex with bound 25(OH) Vitamin D from samples or the Alkaline phosphatase conjugate.

A pNpp substrate solution is added initiating an alkaline phosphatase catalyzed reaction that generates a yellow color in the solution.

Stop solution is added to stop the substrate reaction and the resulting yellow color is read at 405nm. The amount of signal is inversely proportional to the level of 25(OH) Vitamin D in the sample.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

1 product image

  • ELISA - 25(OH) Vitamin D ELISA kit (ab213966), expandable thumbnail

    ELISA - 25(OH) Vitamin D ELISA kit (ab213966)

    25(OH) Vitamin D ELISA Kit (ab213966) Standard Curve

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

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